Spore state discrimination

ABSTRACT

A flow of air including a fungal spore is directed to a collection cartridge. The spore is trapped within the cartridge and positioned within a field of view of a camera sensor. A UV light is activated to illuminate the spore and a camera shutter is opened for a time period. The camera sensor collects light emitted from the spore during a first portion of the time period. After the first portion has elapsed, first and second bursts of white light originating from first and second positions, respectively, are directed towards the spore during a second portion of the time period. After the second portion of the time period has elapsed, the camera shutter is closed to generate an image. The image is analyzed to obtain a shape of the spore.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application claims the benefit of U.S. provisional patentapplication 62/370,604, filed Aug. 3, 2016, and is acontinuation-in-part of U.S. patent application Ser. No. 15/178,170,filed Jun. 9, 2016, which claims the benefit of U.S. provisional patentapplications 62/173,280, filed Jun. 9, 2015, and 62/210,253, filed Aug.26, 2015, and which is a continuation-in-part of U.S. patent applicationSer. No. 15/061,883, filed Mar. 4, 2016, which claims the benefit ofU.S. provisional patent applications 62/129,571, filed Mar. 6, 2015, and62/188,606, filed Jul. 3, 2015, all of which are incorporated byreference along with all other references cited in this application.

BACKGROUND

Minimizing damage from fungal pathogens, such as molds, is ofconsiderable importance to agriculture. There is a need for bettermethods to promptly detect outbreaks of fungal infections, to monitorthe spread of fungal infections, and to track the success of countermeasures such as the application of fungicides.

For example, farms and vineyards can suffer from certain types ofmold—which is a type of fungus—as winds can carry mold spores for manymiles. Depending on climatic conditions, losses for vineyards may rangefrom about 15 percent to about 40 percent or more of the harvest. Theloss in harvest results in lost revenue, profit, and jobs. There is aneed to cost-effectively and rapidly detect damaging mold spores so thatcontrol and mitigation measures can be quickly developed and deployed tosave a harvest.

Agriculture has developed various countermeasures to fungal infectionsof crops, including fungicides. There is interest both in detecting thepresence of a fungal infection as well as monitoring the progress ofanti-fungal countermeasures.

Identifying the state of a fungal spore, such as whether the spore isvirulent or sterile, is important in measuring the success offungicides. There is, however, a lack of real-time systems andtechniques to identify whether or not a fungal spore is virulent orsterile. Further, existing systems and techniques involve very expensiveequipment and are thus out-of-reach for many farmers and vintners. Forexample, techniques using scanning electron microscopy (SEM) oratomic-force microscopy (AFM) are far too time consuming and expensive.When using an optical microscope to view a transparent fungal spore suchas Erysiphe Necator (aka. Powdery mildew) or Botrytis (aka. Gray mold) auser will typically apply a staining dye to enhance the outline of thespore in order to determine the shape (morphology) of the spore;preparing stained spore samples adds undesired cost and delays.Techniques involving the use of fluorescent dyes are also not amenableto automated real-time field measurements. There is a need for improvedsystems and techniques to quickly and cost-effectively determine thestate of a fungal or mold spore.

BRIEF SUMMARY OF THE INVENTION

In an embodiment, there is a method of determining a state of a fungalspore. A flow of air including a fungal spore is directed to acollection cartridge. The fungal spore is trapped within the collectioncartridge. The fungal spore is illuminated with visible light and afirst image of the fungal spore is captured while the fungal spore isilluminated with the visible light. The first image is analyzed toidentify an outline of the fungal spore. The fungal spore is illuminatedwith ultraviolet (UV) light and a second image of the fungal spore iscaptured while the fungal spore is illuminated with the UV light. Ameasurement is made of a degree of fluorescence within the outline ofthe fungal spore. A state of the fungal spore is determined based on thedegree of fluorescence.

In another embodiment, an airborne biological particle monitoring devicecollects particles floating in air. The monitor includes a camerasensor, illumination source, and distinguishes between different states,such as states of virulence of biological particles includingagricultural pathogens. The camera sensor forms part of a highlyintegrated camera sensor chip package including a pixel sensor array,analog drive and readout circuitry, analog-to-digital conversioncircuitry, digital image processing circuitry, and digitalcommunications circuitry.

In another embodiment, a method includes directing a flow of airincluding a fungal spore to a collection cartridge; trapping the fungalspore on a tape medium of the collection cartridge; positioning thefungal spore within a field of view of a camera sensor while the fungalspore remains trapped on the tape medium of the collection cartridge;activating an ultraviolet (UV) light source to illuminate the trappedfungal spore with UV light; opening a camera shutter associated with thecamera sensor for a time period; while the trapped fungal spore isilluminated with the UV light, allowing the camera sensor to collectlight emitted from the trapped fungal during a first portion of the timeperiod; after the first portion of the time period has elapsed,directing, during a second portion of the time period after the firstportion of the time period, a first burst of white light, originatingfrom a first position, towards the trapped fungal spore; directing,during the second portion of the time period, a second burst of whitelight, originating from a second position, different from the firstposition, towards the trapped fungal spore; after the second portion ofthe time period has elapsed, closing the camera shutter to generate animage; and analyzing the image to obtain a shape of the trapped fungalspore.

BRIEF DESCRIPTION OF THE FIGURES

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application with color drawing(s)will be provided by the Office upon request and payment of the necessaryfee.

FIG. 1 shows a block diagram of an airborne particle monitoring systemaccording to an embodiment.

FIG. 2 shows another block diagram of an airborne particle monitoringsystem according to an embodiment.

FIG. 3 shows a block diagram of an airborne particle monitor accordingto one embodiment.

FIG. 4 shows a block diagram of an airborne particle monitor accordingto another embodiment.

FIG. 5 shows a block diagram of a cloud server according to anembodiment.

FIG. 6 shows a block diagram of an airborne particle monitor accordingto another embodiment.

FIG. 7 shows an exterior view of an airborne particle monitor accordingto a specific embodiment.

FIG. 8 shows an isometric view of a particle media cartridge that may beused with the particle monitor shown in FIG. 7.

FIG. 9 shows a plan view of a cross section of the cartridge shown inFIG. 8.

FIG. 10 shows a plan view of a cross section of the particle mediacartridge including media of the cartridge shown in FIG. 8.

FIG. 11 shows a plan-view of the particle monitor shown in FIG. 7including motors.

FIG. 12 shows a vertical cross-section of the particle monitor shown inFIG. 7 illustrating the placement of electronic boards.

FIG. 13 shows some detail of the particle monitor shown in FIG. 7 withoptics and particle media cartridge, as well as illustration of airflow.

FIG. 14A shows a side view of an inside portion of the particle monitorshown in FIG. 7.

FIG. 14B shows a block diagram of a highly integrated camera sensor chippackage.

FIG. 15 shows an enlarged side-view cross-section of the particlemonitor shown in FIG. 7 showing further details of the optical andillumination system according to a specific embodiment.

FIG. 16 is a graph showing spectral characteristics typical of an RGBcamera sensor.

FIG. 17 is a graph showing the absorption spectrum of chlorophyll-a.

FIG. 18 shows a top view of an inspection platform of a particle monitoraccording to another specific embodiment.

FIG. 19 shows a plot combining camera-sensor sub-pixel spectralcharacteristics as shown in FIG. 16 with illumination source spectralcharacteristics.

FIG. 20 shows an overall flow illustrating some basic ingredients ofautomated particle (e.g., mold spore or pollen) monitoring according toa specific embodiment.

FIG. 21A illustrates a detail of interpretation 230 of FIG. 2 involvingdetermination of particle state as well as type.

FIG. 21B shows a block diagram of a particle monitoring device foridentifying a state of a particle according to a specific embodiment.

FIG. 21C shows an example of state determination results that may beoutput according to a specific embodiment.

FIG. 21D shows another example of state determination results that maybe output according to a specific embodiment.

FIG. 22 shows an overall flow of a process for identifying anddetermining a state of a particle according to another specificembodiment.

FIG. 23 is a schematic representation of a fungal spore image usingvisible light illumination.

FIG. 24A is a schematic representation of a healthy fungal sporefluorescence image using ultraviolet light illumination.

FIG. 24B shows an image of a mold particulate matter including sporesfresh off a grape vine.

FIG. 24C shows an image of the mold particulate matter including sporesfifteen hours later.

FIG. 25 is a schematic representation of a fungal spore with a weakfluorescence image using ultraviolet light illumination.

FIG. 26 is a schematic representation of a fungal spore with an unevenfluorescence image using ultraviolet light illumination.

FIG. 27 illustrates mode of action of representative fungicides.

FIG. 28A shows an illumination system of a particle monitor according toa specific embodiment.

FIG. 28B shows an overall flow for combining UV and white light burstingwhen imaging a trapped particle according to a specific embodiment.

FIG. 28C shows an image of a fungal as illuminated under white light.

FIG. 28D shows an image of the fungal as illuminated under UV light.

FIG. 28E shows an image of the fungal as illuminated under UV light anda burst of white light.

FIG. 28F shows further detail of a flow for combining UV and white lightbursting when imaging a trapped particle according to another specificembodiment.

FIG. 28G shows a timeline of events for combining UV and white lightbursting when imaging a trapped particle according to a specificembodiment.

FIG. 28H shows an image generated where UV light was coming from abottom right hand corner of the field of view.

FIG. 28I shows an image generated where a direction of illumination wasfrom a bottom left hand corner of the field of view.

FIG. 28J shows an image generated where a direction of illumination wasfrom a top left hand corner of the field of view.

FIG. 28K shows an image generated where a direction of illumination wasfrom a top right hand corner of the field of view.

FIG. 28L shows an image generated where all three white LEDs were onsimultaneously for 1 second.

FIG. 28M shows an image generated where illumination included a singlewhite LED and no UV light was present.

FIG. 28N shows an image where the white light bursting or flashingoccurred early during the exposure time period.

FIG. 28O shows an image where the white light bursting or flashingoccurred late during the exposure time period.

FIG. 28P shows a top view of an illumination system of a particlemonitor according to another specific embodiment.

FIG. 28Q shows a top view of an illumination system of a particlemonitor having a movable light source in a first position according toanother specific embodiment.

FIG. 28R shows a top view of an illumination system of a particlemonitor having a movable light source in a second first positionaccording to another specific embodiment.

FIG. 28S shows a graph of fluorescence color characteristics forriboflavin, NADH, and tryptophan.

FIG. 29 shows corresponding biomolecule fluorescence wavelengthconversions for the color characteristics shown in FIG. 28S.

FIG. 30 shows a formula for relating measured pixel values“RGB_(i)(x,y)” to the biomolecule distributions “C_(j)(x,y).”

FIG. 31 shows the application of the formula shown in FIG. 30 to thebiomolecule fluorescences of FIG. 29.

FIG. 32 shows a flow chart of a process for assessing a fungal sporeaccording to a specific embodiment.

FIG. 33 shows another flow chart of a process for assessing a fungalspore according to a specific embodiment.

FIG. 34 shows a block diagram of a particle information packet accordingto an embodiment.

FIG. 35 shows a block diagram of history for the particle informationpacket according to an embodiment.

FIG. 36 shows a deployment of a prototype particle monitor in avineyard.

FIG. 37 shows an external solar panel and battery used to power theprototype particle monitor shown in FIG. 36.

FIG. 38 shows a first representation of planar benzene molecule C₆H₆.

FIG. 39 shows a second representation of planar benzene molecule C₆H₆.

FIG. 40 shows a third representation of planar benzene molecule C₆H₆.

FIG. 41 shows a molecular structure of anthracene.

FIG. 42 shows a molecular structure of triphenylene.

FIG. 43 shows a molecular structure of coronene.

FIG. 44 shows a plot of two size distributions of diesel particulatematter (DPM).

FIG. 45 shows another plot of two size distributions of DPM.

FIG. 46 shows a molecular structure of riboflavin.

FIG. 47 shows a molecular structure of nicotinamide adeninedinucleotide.

FIG. 48 shows a molecular structure of tryptophan.

FIG. 49 shows a section of an air filter cartridge according to aspecific embodiment.

FIG. 50 shows a section of another air filter cartridge according toanother specific embodiment.

FIG. 51 shows a section of a particular monitoring device for airbornediesel soot detection according to a specific embodiment.

FIG. 52 shows a block diagram of a client-server system and network inwhich an embodiment of the system may be implemented.

FIG. 53 shows a system block diagram of a client or server computersystem shown in FIG. 52.

DETAILED DESCRIPTION

FIG. 1 is a block diagram of an airborne particle collection, detectionand recognition system 100 according to a specific embodiment. Thissystem addresses unmet needs of the agricultural industry includingvineyards for customized and actionable information regarding theircrops' exposure to airborne particles such as mold spores, or otherairborne particles. An airborne particle monitoring system asillustrated in FIG. 1 addresses needs related to monitoring of airquality as well as monitoring of airborne agricultural pathogens. Thelatter application is of particular interest to vineyards, farms, andthe agricultural industry.

In the example shown in FIG. 1, the system includes an airborne particlemonitoring device 105 and a remote cloud server 110 that is connected tothe monitoring device via a communication network 115. Theparticle-monitoring device may be referred to as a particle detector,spore detector, pollen detector, particle collector, spore monitor,pollen monitor, spore collection machine, pollen collection machine, orairborne biological particle monitoring device.

The monitoring device is a device or appliance that is designed to beplaced in a local environment 120 where monitoring is desired. In aspecific embodiment, the monitoring device is contained within acylindrical housing having a diameter of about 100 millimeters (mm) anda height of about 150 mm. The device may be placed outdoors inagricultural fields, in vineyards, or other locations where it is ofinterest to monitor the presence and state of agricultural pathogenssuch as fungal spores.

Information including alerts and notifications can be sent from theparticle monitor, cloud server, or both to a client (e.g., mobile)device 135 of a user 125. The user may be, for example, an agriculturaldisease management consultant or the owner or manager of a vineyard orfarm. In an embodiment, the user can use their mobile device to sendinstructions or commands to the particle monitor, receive notificationsand alerts from the particle monitor, or both. In an embodiment, theparticle monitor is a network-enabled device. The user (and their mobiledevice) can be remote from the particle monitor. For example, theparticle monitor may be placed in a vineyard in Napa, Calif. and theuser may be in New York, N.Y. The particle monitor can sendnotifications, alerts, and exchange communications with the mobiledevice via the network.

An agricultural operation may be susceptible to known pathogens such aspowdery mildew. The system shown in FIG. 1 can be used to help identifythe specific types of airborne particles such as powdery mildew spores.With this information, a plan for countermeasures can be developed toreduce or eliminate crop infection from agricultural pathogens such as aplan for the targeted application of selected fungicides. In a specificembodiment, the monitoring device samples ambient air 130 and collectsor traps airborne particles that may be present or floating in theambient air.

The monitoring device can use a combination of techniques to analyze,discriminate, and identify the collected particles. In a specificembodiment, the analysis includes capturing images (e.g., pictures,photographs, or snapshots) of the particles under various lightingconditions and examining the captured images. Particles, includingdifferent types of spores and pollen, can be identified or discriminatedbased on their morphology (e.g., shape, surface texture, apertures, orsize), color, fluorescence characteristics, or combinations of these asmay be captured by a camera sensor of the particle monitor.

In another specific embodiment, the analysis further includes combiningthe image analysis with context information that is obtained from theremote cloud server. The context information may include, for example,information regarding weather, wind conditions, humidity levels, thetypes of spores and pollen currently propagating at the geographicallocation of the collected particles, vegetation known to be present atthe geographical location of the collected particles, other contextinformation, or combinations of these.

For example, in a specific embodiment, a particle monitoring devicegenerates a set of candidate particle identifications for a particularparticle that has been captured based on analyzing a set of images takenof the captured particle. After the set of candidate identificationshave been generated, the particle monitoring device issues a request tothe cloud server for context information. The request can include ageographical location of the particle monitoring device, time and dateof particle capture, or both. The cloud server receives the request anduses the geographical location of the monitoring device, time and dateof capture, or both to retrieve appropriate or relevant contextinformation to transmit to the monitoring device.

The monitoring device receives the appropriate context information andfurther analyzes the context information in conjunction with the set ofcandidate particle identifications. Consider, as an example, thatparticles are identified as powdery mildew spores, but by itself themonitoring device cannot identify which of several species of powderymildew has been detected. If, however, the context information receivedby the particle monitor from the cloud server indicates that only one ofthe many species of powdery mildew is currently propagating inagricultural fields at the geographical location of the monitoringdevice, the analysis may conclude that the species detected is thespecies known to be propagating in the geographical area.

The images of the particles captured by the monitoring device may betransmitted or sent to the remote cloud server for further analysis. Theanalysis may include a review of the images by a human technician. Forexample, in some cases, an automated image analysis and contextinformation analysis may not lead to a satisfactory identification. Inthese cases, the analysis may be escalated to a human technician. Inparticular, the images, associated metadata, or both can be transmittedto the cloud server for review by a human technician. The associatedmetadata can include the geographical location of the particle monitor,time and date of particle capture, or both.

In a specific embodiment, the particle monitoring device traps theairborne particles on a piece of media or medium that can be removed bythe user from the particle monitoring device. The media, with trappedairborne particles, may additionally be transported to a lab for anin-depth analysis. Consider, as an example, that the human technician isunable to identify with reasonable certainty the particle from theimages. The technician can escalate the analysis to an analysis of theactual collected particle. In particular, the technician can notify theuser that the collection media should be removed from the particlemonitoring device and delivered to a laboratory for an analysis of theactual collected physical particles. For example, the technician maytransmit through the system a notification to an app executing on theuser's mobile device. The app may display the message, “Please removeparticle collection media from your particle monitor and deliver it toour laboratory for analysis.”

This technique of escalation is an example of what may be referred to astiered particle analysis. Such an analysis helps to ensure judicious useof resources including computing resources (e.g., network bandwidth,storage) and human labor. Activities such as accessing a network,sending image files over the network, human review, delivering thephysical particles, and so forth consume resources. For example, animage file may be several megabytes in size. It can be desirable torefrain from transmitting the image file over a network unless thetransmission is deemed necessary.

In a specific embodiment, there is a first attempt to identify thecollected particles where the first attempt is performed locally (e.g.,at the particle monitor). If the first attempt fails to result in asatisfactory identification, a second attempt includes accessing aremote cloud server to obtain over a network context information. If thesecond attempt fails to result in the satisfactory identification, athird attempt includes transmitting over the network the image files tothe remote cloud server for human review. If the third attempt fails toresult in the satisfactory identification, a fourth attempt includesinstructing the user to mail the removable media with collectedparticles to a laboratory.

In a specific embodiment, the particle monitoring device is paired withone or more mobile devices 135 associated with or belonging to the usersuch as an agricultural disease management consultant. The pairingallows the particle monitoring device and mobile device to exchangeinformation, instructions, data, commands, or other communications.

Mobile devices include, for example, smartphones, tablet computers, andwearable computers (e.g., Apple Watch, Google Glass). Mobile devices 135are not limited to consumer products and also may include airbornedrones and automated land-based vehicles. For example, a number ofparticle monitoring devices may be located at various positions within acultivated field. An airborne drone may do a survey of the cultivatedfield and detect a possible issue in the vicinity of one of the particlemonitoring devices. The drone may then fly in close proximity to thatspecific particle monitoring device and wirelessly collect data that hasaccumulated in the particle monitoring device.

Conversely, a particle monitoring device may detect a pathogen ofconcern and activate a beacon signal that causes a drone to fly by tocollect data accumulated in the particle-monitoring device as well as tosurvey the portion of the cultivated field in proximity to theparticle-monitor device for possible signs of crop damage. Similarly,particle monitoring devices may interact with automated land-basedvehicles. Both airborne drones and autonomous land-based vehicles may beequipped with sprayers and may be programmed to spray fungicides in thevicinity of particle-monitoring devices that have detected a level ofpathogenic spores exceeding some preprogrammed threshold. Mobile devices135 may be any generalized to any interacting set of mobile devices,fixed devices, or both that are in communication with one or moreparticle monitoring devices.

In a specific embodiment, a method includes sending data to airbornedrones, automated land-based vehicles, or both. There can be a dronethat does a survey of land and detects possible issues and wants to geta read of particle readings, or vice versa. That is, the particlemonitoring device may detect continuous high particulate in the vicinityand may send data to a drone that will then perform a survey of a largerarea.

Similarly, data could be sent to an autonomous land-based sprayer thatmay decide to come in a spray based on some preprogrammed thresholds ofparticles over time. In one embodiment, a method includes deploying oneor more particle monitors to monitor a cultivated field, configuring fora particle monitor a threshold including a number pathogen detectionsover a period of time, detecting, at the particle monitor, a set ofpathogens of concern over the period of time, and if a number of the setof pathogens of concern exceeds the threshold, activating, at theparticle monitor, a beacon signal, where the beacon signal is to bereceived by an autonomous vehicle.

The autonomous vehicle may include a tank holding a pesticide, and asprayer to spray at least a portion of the cultivated field with thepesticide upon receipt of the beacon signal by the autonomous vehicle.The autonomous vehicle may include an airborne drone configured tosurvey at least a portion of the cultivated field upon receipt of thebeacon signal by the autonomous vehicle. The autonomous vehicle mayinclude an airborne drone. The autonomous vehicle may include a landvehicle.

In another specific embodiment, a method includes deploying one or moreparticle monitors to monitor a cultivated field, receiving at a particlemonitor a command from an autonomous vehicle, and upon receipt of thecommand from the autonomous vehicle, sampling by the particle monitorambient air.

FIG. 2 is a block diagram showing some elements and processes of anairborne particle collection, detection, and recognition system 200according to a specific embodiment. Airborne particle monitoring system200 includes air intake hardware 220 to sample ambient air 210,particle-capture hardware 222 with which particles removed from ambientare collected and transported for microscopic analysis. The microscopesystem includes illumination hardware 224 that shines visible,ultraviolet (UV), or infrared (IR) light, or combinations of these oncaptured particles, and image-capture hardware 226 that may include alens assembly as well as a camera sensor. The light may include lightemitted from quantum dots. Capturing various images of the particleswhen illuminated under different conditions provides for additionaldimensions of analysis to help identify, classify, or discriminatebetween the collected particles.

Image processing software and hardware 228 processes image data from theimage-capture hardware 226. The types of the observed particles are thendecided by interpretation software 230. Finally, user-notificationsoftware 232 outputs the interpretation results in a form that can beunderstood by the user. For example, the output may include displayingon an electronic screen a message specifying the airborne particles thathave been collected and identified. The value of airborne particlemonitoring system 200 can be realized when it beneficially guides theuser to take an informed user action 280.

In some embodiments, particle-capture hardware 222 provides for a mediumthat can be removed with captured particles and archived for possiblefuture laboratory inspection, thus providing a physical archive 260 ofcaptured particles.

The actions and data processing of airborne particle monitoring system200 is orchestrated through a local processor 240. Local processor 240is preferably supported by other computing and data resources viadigital communication networks that may concisely be referred to as the“cloud”; see, e.g., cloud server of FIG. 1.

Local processor 240 and cloud 250 may support numerous feedback loops.Here is one example. Interpretation software 230 (which may be codeexecuted in a dedicated processor, or by the local processor, or on thecloud) may be unable to reach a definitive result and the system mayrespond by requesting ultraviolet light illumination from theillumination hardware 224 in order to generate additional fluorescencespectral information.

In an embodiment, the image capture hardware 226 is based on an imagingsensor designed for use in color cameras. The mass market for digitalcameras, including those in smartphones, has resulted in very capablecolor camera sensors at relatively low prices. Such color camerasensors, such as the SON-IMX028 CMOS image sensor by Sony, provides atlow cost rich data for particle detection and discrimination.Furthermore, the spectral richness of data collected with a color camerasensor may be extended by enhancing the capabilities of the illuminationhardware 224; more details are given further below. The use of colorcamera sensors in combination with enhanced illumination hardware isadvantageous for the goal of providing a capable airborne particlemonitoring system 200 in a competitive price.

It is of interest to note that the low-cost of such color camera sensorsis not just a matter of high manufacturing volumes, but also the highdegree of integration of the products. One example of a highlyintegrated and low-cost color camera sensor suitable for use with theparticle monitor is SONY's SON-IMX028 CMOS image sensor as provided bySony Corporation of Tokyo, Japan. Such highly-integrated RGB camerasensors include within their chip package not only the RGB pixel sensoritself, but also associated analog drive and readout circuitry,analog-to-digital conversion circuitry, digital circuitry for imagecapture and processing as well as digital electronics sufficient tooutput images in a digital format to an external main processor.

FIG. 3 shows a block diagram of particle monitoring device 105 accordingto one embodiment. The block diagram shows a number of subsystems,processing modules, and components that can be included in the particlemonitoring device. In an embodiment, the particle monitoring deviceincludes quantum dots. The quantum dots are configured emit light havingspectral characteristics corresponding to a particle of interest.Particles collected by the monitoring device are illuminated under thelight emitted by the quantum dots and a color image (e.g., picture,photograph, or snapshot) is taken while the particles are illuminated bythe quantum dots. The image can be analyzed to determine whether theimage includes the particle of interest.

The particle monitoring device shown in FIG. 3 includes a device housingor enclosure 302 and a base 304, connected to the housing. The housingincludes an air intake opening 306 and an air exhaust opening 308.Contained within or inside the housing is a blower 314, removableparticle collection media 316, collection media motor 318, illuminationsubsystem 320, optical subsystem 322, particle identification subsystem324, display 326 connected to the particle identification subsystem,storage 328, network interface controller 330, Bluetooth communicationcard 332, global positioning system (GPS) sensor 333, housing motor 334,and power source (e.g., battery) 336. There can be a user-operable powerswitch so that the device can be turned on, turned off, placed in astandby state, or combinations of these.

The subsystems, processing modules, components and so forth areconnected by a bus system 338. The power source provides electricalpower to the subsystems, processing modules, and components. The powercan be DC power such as that provided by a battery. Using a battery tosupply power facilitates a particle monitor that may be placed in anoutdoor environment such as an agricultural field. A particle monitormay include a set of solar cells for recharging the battery. In anotherspecific embodiment, the power can be AC power.

The blower is responsible for moving ambient air outside the monitordevice housing, through the air intake opening, into the monitor devicehousing, towards the collection media, and then out through the airexhaust opening. The blower may be referred to a fan.

The removable particle collection media provides a medium for trappingparticles that are airborne or floating in the ambient air. In aspecific embodiment, the collection media includes an adhesive tape. Thetape is flexible so that it can be mounted on or wound upon on a reel orspool. The adhesive tape includes a backing material and an adhesivethat is applied to a side of the backing material. The backing materialcan be made of paper, plastic, plastic film, fabric, polyester, Teflon,nylon, cloth, metal foil, or any other competent material. The adhesivecan be any type of adhesive that can trap particles floating in theambient air. The adhesive may include glue, paste, mastic, rubbercement, or other sticky or tacky substance. The blower directs the flowof air towards or over the collection media. Particles within the airare then trapped by the adhesive-coated side of the tape.

In a specific embodiment, the tape is 3M polyester film tape 850 asprovided by 3M Corporation Maplewood, Minn. Applicants have discoveredthat this particular tape includes features desirable for a particlemonitor. In particular, the polyester film includes a wide temperaturerange resistance (e.g., −50 degrees Celsius to 177 degrees Celsius)which helps to reduce failure caused by film shrinkage or embrittlement.The wide temperature range resistance is desirable because in someembodiments, the monitor device is used outdoors and thus must survivewide temperature fluctuations throughout the day and times of the year.For example, temperatures typically drop during the night and riseduring the day. Applicants have discovered that for applications ofparticle monitoring the tape shows desirable, long lasting resistance tocyclic fatigue. This means the tape can be pulled off and coiled againmultiple times to re-examine trapped particles again and again and thetape still retains very good adhesion.

In a specific embodiment, the adhesive on the tape includes an acrylicadhesive. This is advantageous because it is not water-based and thuscan better survive outdoor environments. For example, outdoorenvironments can be more subject to moisture as compared to indoorenvironments. An acrylic adhesive can tolerate moisture better than awater-based adhesive. In a specific embodiment, the tape includes apolyester film. Properties desirable in the polyester film—including itswide temperature range—is that it can be made very thin, possesses veryhigh strength, has high moisture resistance, and is resistant tochemicals and solvents (e.g., will not decompose easily if chemicals orsolvents floating in the air should fall on the tape).

It should be appreciated that 3M polyester film tape 850 is merely oneexample of a tape suitable for use with the particle monitor and inother embodiments, other tapes with properties desirable for theparticle monitor may instead be used. For example, 3M film tape 850includes an adhesion to steel specification according to ASTM testmethod D-3330 of 31.5 N/100 mm. The collection media motor is designedwith sufficient power to advance and uncoil the tape. Applicants havefound that a lower adhesion to steel value can be desirable (e.g., about15.7 N/100 mm) because less power is required to advance and uncoil thetape.

In a specific embodiment, a color of the tape is black or a dark color.An advantage of using black or a dark color is that light is less likelyto reflect or bounce off the tape as compared to lighter colors (e.g.,white). For example, a technique of the system includes capturing imagesof the particles under different specified illumination conditions.Light (e.g., white light) bouncing off the tape and into the camerasensor may skew the images and, in particular, the colors captured inthe images. In another specific embodiment, the tape is transparent orat least partially transparent. A transparent tape allows for backsideillumination (e.g., illuminating from below the tape).

In a specific embodiment, a removable cartridge is provided which housesthe adhesive coated tape. The cartridge houses a supply reel, an uptakereel, and the adhesive coated tape. An end of the tape is connected tothe supply reel. An opposite end of the tape is connected to the uptakereel. The adhesive coated tape is wound upon the supply reel and spentportions of the tape upon which particles have been trapped are woundonto the uptake reel. The cartridge may further include anidentification tag such as a radio frequency identification tag (RFID)tag, machine-readable code (e.g., barcode, quick response (QR) code), orother label. Depending upon the type of tag, the tag may be attached toa body of the cartridge (e.g., via glue), or printed onto the body ofthe cartridge. The particle monitor may include a corresponding reader.The identification tag allows the particle monitor to uniquely identifythe cartridge.

In another specific embodiment, the collection media includes a rigiddisc. A side of the disc is coated with an adhesive to trap the airborneparticles that enter the monitoring device. The disc exposes differentregions around an annulus so that particles are trapped within aparticular region. The disc may be made of plastic, nylon, metal, or anyother rigid material. In another specific embodiment, the collectionmedia includes adhesive-coated glass slides. In each embodiment, theadhesive coated tape (or other particle collection media such asadhesive-coated glass slides or adhesive-coated disc) may be removedfrom the particle collection device and fresh media inserted into theparticle collection device. Anywhere a glass slide may be used, aplastic slide is likely to be an equally viable option. Removed mediacontaining captured particles may be subjected to laboratory inspectionand testing, archived for possible future laboratory inspection andtesting, or both.

The collection media motor is responsible for advancing the collectionmedia. For example, in an embodiment, the collection media includes acartridge having a supply reel, an uptake reel, and an adhesive coatedtape wound about the supply reel and connected to the uptake reel. Uponcollecting some airborne particles on a portion of the adhesive coatedtape, the media motor can advance the tape so that new particles can betrapped on another portion of the adhesive coated tape. The portionhaving the previously trapped airborne particles can be advanced to theparticle identification subsystem for imaging and examination.

The collection media motor may include a counter that tracks a positionof the tape. The position of the tape can be associated with the image.Storing the position information allows the tape to be later advanced(or unwound) to the same position at which the image was taken andadditional analyses to be performed. The counter may count a number ofunits between a reference point on the tape (e.g., a beginning of thetape or an ending of the tape) and a location of the tape at which theimage was taken. The units may be distance-based. For example, thelocation of the tape may be a distance as measured from the beginning ofthe tape.

The illumination subsystem includes various optical elements forgenerating and emitting light or radiation (e.g., visible light,ultraviolet light, infrared, or combinations of these) into theparticles that have collected on the collection media. The illuminationsubsystem includes one or more light sources (e.g., two light sources).Each light source includes one or more light emitting elements.

In a specific embodiment, a lighting element includes a light emittingdiode (LED). A light source may include a cluster of light emittingelements such as a cluster of LEDs (e.g., two or more LEDs). A clustermay include any number of light emitting elements such as LEDs. Forexample, a cluster may include one, two, three, four, five, six, seven,eight, or more than eight LEDs. In another specific embodiment, alighting element includes a laser diode. There can be a combination ofdifferent types of light emitting elements such as a combination of LEDsand lasers.

The illumination subsystem may include lenses, filters, diffusers, orcombinations of these for directing or modifying the light as desired.For example, a diffuser may be used to spread out the light from alighting element and provide a soft light. A diffuser can help to ensurethat the area around the collected particles is illuminated. In aspecific embodiment, the illumination system includes optical fiber. Theoptical fiber can be used to collect light emitted by a light source anddirect the light onto the collected particles. Optionally, theillumination system may also include polarizers or light sources thatare inherently polarized.

In an embodiment, the illumination subsystem includes a first lightsource 344, and a second light source 346. In an embodiment, at leastone of the first or second light sources is an ultraviolet light source(e.g., radiation wavelengths ranging from about 10 nm to about 380 nm).For example, an ultraviolet light source may include an LED with acharacteristic emission wavelength of 365 nm. This is a relatively longultraviolet wavelength with sufficient photon energy to excitefluorescence in flavin molecules but few other biomolecules. Additionalultraviolet light sources of shorter wavelengths and higher photonenergies may be provided that induce fluorescence in additionalbiomolecules.

The optical subsystem includes various optical elements for capturingone or more images of the collected particles while the collectedparticles are being illuminated or radiated by the illuminationsubsystem. In an embodiment, the optical subsystem includes a microscopeincluding a camera sensor 348 and lens assembly 350. A microscope is anoptical instrument having a magnifying lens or a combination of lensesfor inspecting objects too small to be seen or too small to be seendistinctly and in detail by the unaided eye. The lens assembly includesa set of lenses for bringing the collected particles into focus,magnifying the collected particles, or both. The camera sensor collectslight scattered or reflected, or fluorescently re-emitted, back from theparticles to capture images or photographs. Optionally the opticalsubsystem may include one or more polarizers so that images may becaptured for light of known polarization.

The particle identification subsystem includes an image recognitionengine 352, particle reference library 354, and context informationacquisition unit 356. A particle identification manager 358 manages theparticle identification or discrimination process.

The particle reference library stores reference information identifyingdifferent types of airborne particles. In a specific embodiment, thereference information includes particle-discrimination algorithmparameters. Optionally, these particle-discrimination algorithmparameters are determined by machine learning algorithms and a learningset of reference files that includes images including color photographs,fluorescence images, or both of different types of known particles. Themachine learning algorithms that determine the particle-discriminationalgorithm parameters may run locally, on the cloud, or both. Running thealgorithms in the cloud helps to reduce the cost of computing hardwarein the local device. Being able to run the algorithms locally at theparticle monitor can be advantageous in environments where there islimited network connectivity. The set of learning files may includereference images of mold spores, pollen, and other particles ofinterest. Table A below shows an example of a data structure that may beused to store the reference information.

TABLE A Filename Description powdery_mildew_spores.jpg Picture ofpowdery mildew spores. Botrytis_spores.jpg Picture botrytis spores. . .. . . .

A first column of the table is labeled filename and lists the variousfiles stored in the particle reference library. A second column of thetable includes metadata (e.g., a description) that identifies the objectin the corresponding file.

In an embodiment, the image recognition engine receives the image of thecollected particles taken by the optical subsystem and analyzes theimage using particle-discrimination algorithm parameters it previouslyreceived from the cloud. For example, particle-discrimination algorithmsrunning in the particle identification subsystem may identify thecollected particle as a spore of the pathological vineyard mold sporebotrytis.

Some examples of parameters that may be considered in aparticle-discrimination algorithm include autofluorescence properties(e.g., intensity of autofluorescence), size, shape, length of polaraxes, length of equatorial axes (or diameter), ratio of polar axis toequatorial axis (P/E ratio), number of apertures, type of apertures,shape of apertures, position of apertures, lack of apertures, colorcharacteristics, geometrical features, type of symmetry (e.g., radialsymmetry or bilateral symmetry), lack of symmetry, other parameters,weights, or combinations of these. One or more of these parameters maybe derived or extracted from optical system measurements, specified as athreshold, and then used as a discrimination algorithm parameter todiscriminate particles.

The image recognition engine may use any competent technique orcombination of techniques for recognizing the particles imaged by theoptical subsystem. Some examples of image recognition techniques includeedge detection, edge matching, changes in color, changes in size,changes in shape, divide-and-conquer searches, greyscale matching,gradient matching, histograms of receptive field responses, large modelbases, interpretation trees, hypothesize and test, post consistency,pose clustering, invariance, geometric hashing, scale-invariant featuretransform (SIFT), and speeded up robust features (SURF), among others.

The context information acquisition unit is responsible for obtainingcontext information associated with the particles that have beencollected by the monitoring device. The context information may be basedon a geographical location of the collected particles, a time and dateof the collection, or both. In an embodiment, the context informationincludes known agricultural pathogens recently detected in one or morevarious particular geographical areas. For example, if a short list ofspecies of powdery mildew is known to infect vineyards in Napa Valley,then a particle monitor located in a vineyard in Napa Valley may beprovided with this list. The context information may include weatherconditions, temperature, wind speed, wind patterns, and so forth.

The context information may include a listing of particle types thathave been identified by other nearby particle monitors, mobile drones,or both. For example, nearby particle monitors may include particlemonitors that are within a specified radius of the requesting particlemonitor. The radius may be, for example, 50, 100, 500, 1000, 2000, ormore than 2000 meters. The radius may be less than 50 meters. The radiusmay be configurable such as by a user or administrative user. The radiusmay be determined dynamically. For example, the radius may varyproportionally to current wind speed as high winds can increase thelikelihood of particles being carried into the local environment fromremote areas.

The context information is used by the particle identification subsystemto help narrow the list of candidate particle types. Results of theparticle identification subsystem may be outputted to the display,recorded in a log, or both.

The storage may include a particle identification log 360, imagesrepository 362, and image index 364. The particle identification logrecords identifications of particles as determined by the particleidentification subsystem. Table B below shows an example of informationthat may be recorded in the log.

TABLE B Image Context Info Particles File File Present TimestampLocation 001.jpg Contextl.txt Grass Apr. 10, 2016, 45 Appleseed Drive,pollen 11:34 AM Santa Rosa, CA 94555 002.jpg Context2.txt Grass Apr. 10,2016, 45 Appleseed Drive, pollen 1:36 PM Santa Rosa, CA 94555 003.jpgContext3.txt Botrytis Apr. 11, 2016, 45 Appleseed Drive, spores 2:00 PMSanta Rosa, CA 94555

In the example shown in table B above, a first column of the table liststhe name of the file containing the image of the collected particles. Asecond column lists the name of the file containing the contextinformation that may be associated with a geographical location of thecollected particles, time and date of the collected particles, or both.The context information may be formatted as a text file, ExtensibleMarkup Language (XML) formatted file, or in any other file format asdesired. A third column identifies type(s) of particle(s) of interest. Afourth column of the table stores a timestamp indicating a time and datethat the particles were collected. A fifth column of the table stores alocation of the particle collection.

It should be appreciated that the data shown in table B above is merelyan example of some of the metadata information associated with particleidentification that may be stored in the database. In a specificembodiment, a particle information packet and particle informationpacket history is stored. Further details are provided below.

The images repository stores the image files generated by the opticalsubsystem. The files store digital images of the particles that havebeen captured. The files may include raw image files (e.g., digitalnegatives), raster images, bitmapped images, or combinations of these.The files may be formatted using any type of image file format (e.g.,jpeg, exif, tiff, gif, bmp, png, and so forth).

The image index database stores metadata associated with the imagefiles. The metadata may include, for example, image filenames, time anddate that the image was taken, geographical location data, opticalsettings, and so forth. The metadata may include a description orspecification of the lighting conditions, as provided by theillumination subsystem, under which the images were made. For example,the metadata may indicate that a first image was taken while particleswere illuminated by white light, a second image was taken while theparticles were illuminated by red light emitted from quantum dots, athird image was taken while the particles were illuminated byultraviolet light, a fourth image was taken while the particles wereilluminated by infrared light, and so forth. The index can be accessedand searched.

In a specific embodiment, the particle identification log, particleimage files, image index, or combinations of these are transmitted fromthe particle monitor to the cloud server for further review, archivalstorage, backup. For example, the particle image files may betransmitted to the cloud server periodically or in batch such asnightly, weekly, or at any other frequency or time as desired. Once theimage files have been transmitted to the cloud server, the image filesmay be deleted from the particle monitoring device. Deleting the imagesfrom the particle monitoring device frees up storage space for newimages.

The GPS sensor provides geographical location information. Thegeographical location information allows the images of the collectedparticles to be tagged with the location of collection. As discussed,the location information is used to obtain context information such asfungal species currently propagating at the geographical location ofcollection, weather conditions, identify other nearby particle monitors,or combinations of these.

The Bluetooth communication card or chip allows for a wireless pairingof the particle monitor and a user's mobile device. Bluetooth includes acommunication protocol that allows for communicating over shortdistances (e.g., about 10 meters). The wireless pairing allows theparticle monitor device and mobile device to exchange communication andother information. For example, in a specific embodiment, the particlemonitor transmits to the mobile device a message including anidentification of a particle that was collected. It should beappreciated that Bluetooth is merely one example of a standard forwireless communication. Other embodiments may include othercommunication standards in addition to or instead of Bluetooth such asWiFi. The particle monitor may include a radio transmitter and antennafor long distance communication.

A power subsystem of the particle monitor may include a low-batteryindicator unit. When the available battery power drops below a threshold(e.g., 20 percent battery remaining), the low-battery indicator unit cantransmit a notification such as text message notification to the user'smobile device to notify the user that the particle monitor should berecharged.

The housing motor turns or rotates the particle device housing about thebase. The turning allows the air intake opening to pull in ambient airfrom different directions so that there is a good or representativesampling of air. The housing motor can be used to ensure that the airintake openings are aligned with a direction of wind so that airborneparticles in the wind will enter through the air intake opening.

In a specific embodiment, the power source includes one or morebatteries. The battery may be a rechargeable battery. Examples ofrechargeable batteries include nickel cadmium (NiCd) batteries, nickelmetal hydride (NiMH) batters, lithium ion (Li-ion) batteries, andothers. When the rechargeable battery within the particle monitor isdepleted, the batteries may be recharged by an AC adapter and cord thatmay be connected to the particle monitor. Alternatively, batteries maybe recharged with energy from solar panels. In other words, in anembodiment, the particle monitor does not necessarily require AC powerto recharge. In this embodiment, the particle monitor device may bepowered in the field using solar panels and a rechargeable lead-acidbattery. In this specific embodiment, there can be a controller thatregulates the solar panels load into the battery and into the device.

Instead or additionally, the particle monitor may include a universalserial bus (USB) port. The USB port allows the particle monitor to beconnected to a computer such as a desktop computer for charging. Theport may also be used to configure the particle monitor via the desktopcomputer, transfer data from the particle monitor to the desktopcomputer, transfer data from the desktop computer to the particlemonitor, or combinations of these. In another specific embodiment, thepower source includes one or more disposable batteries.

The network interface controller provides the gateway to communicatewith the mobile device, server, or both. In an embodiment, the networkinterface is connected to the Internet. The network interface controllermay include an antenna for wireless communication, an Ethernet port toconnect to a network via a cable, or both.

The housing may be made from a material such as plastic, nylon, metal,wood, or combinations of these. In a specific embodiment, the housing ismade of plastic. A non-conductive material such as plastic is desirablebecause a plastic housing allows for the passage of radio waves so thatthe particle monitor can communicate wirelessly. For example, an antennalocated inside a plastic housing will be able to receive and transmitwireless signals through the plastic housing. Plastic is also relativelyinexpensive to form and manufacture. In other cases, however, a metalhousing may be desired. Metal can be less likely to crack as compared toplastic and users may prefer the aesthetic appearance of metal. Inembodiments where the housing is made of metal, the antenna may belocated or embedded on an outside surface of the housing.

FIG. 4 shows another specific embodiment of particle monitor 105. Theparticle monitor shown in FIG. 4 is similar to the particle monitorshown in FIG. 3. The particle monitor shown in FIG. 4, however, includesa removable particle collection media with quantum dots 405. In aspecific embodiment, the collection media is contained within acartridge. The cartridge includes a pair of spools and a tape woundabout the pair of spools. The tape includes an adhesive and a backingmaterial where the adhesive has been applied to a side of the backingmaterial. Further detail is provided below.

FIG. 5 shows a block diagram of a remote cloud server 505 according to aspecific embodiment. The server includes a particle identificationupdate module 510, reference library update module 515, contextinformation processing unit 520, central particle image repository 525,central particle identification log repository 530, context informationdatabase 535, database 540 storing information about various registeredparticle monitors that have been deployed, and particle identificationserver engine 545. A technician console 550 is connected to the server.

The particle identification update module is responsible for sendingcode updates to the various particle monitors that have been deployedthroughout the world. The code updates may include firmware updates. Theupdates help to ensure that each monitor is equipped with the mostrecent versions of the algorithms for particle identifications.

The reference library update module is responsible for sending new orupdated reference images of particles. For example, as new referenceimages of particles are made, these reference images can be distributedto each of the various particle monitors. Alternatively, or in addition,the reference library information includes particle-discriminationalgorithm parameters that may be distributed to each of the variousparticle monitors. Storing particle discrimination algorithm parameterscan require less storage space than the reference images.

The context information database stores context information such asclimatic conditions associated with different types of pathogenic fungalspores, blooming periods of various plants and flowers, geographiclocation data for the various plants and flowers, weather conditions,and so forth. The context processing unit can receive from a particlemonitor a request for context information where the request specifies ageographical location of the particle monitor, time of particlecollection, or both. The context processing unit can access the contextinformation database to retrieve a subset of relevant contextinformation corresponding to the geographical location, time, or bothand transmit the subset of relevant context information to therequesting particle monitor.

The central particle image repository stores images of particles thathave been taken by the various particle monitors and transmitted to thecloud server. The images can be accessed and viewed via the technicianconsole by a human technician 555. The central image repository (orother central repository) may further store the analysis results fromthe various particle monitors. This allows the technician to performmanual spot checks of the analysis to help ensure that the particleidentifications made by the particle monitors are accurate. The imagerepository further allows the technician make a manual identification ofparticles by reviewing images where the local particle monitor is unableto make a satisfactory identification.

The central particle log repository stores particle identification logsgenerated by the various particle monitors and transmitted to the cloudserver. As discussed, the particle identification logs can includelistings of particle types that have been identified and associatedmetadata such as a time and date of particle capture, location ofparticle capture, and so forth.

The deployed monitors database stores information about the variousparticle monitors that have been deployed throughout the world. Thedatabase may be referred to as a particle monitor registration database.The information may include, for example, a geographical location of aparticle monitor, particle identification logs containing informationabout particles captured by the particle monitor, images or an index toimages taken by the particle monitor, user information (e.g., companyname, name of primary contact, email address, or mailing address) dateparticle monitor was purchased, device serial number, firmware version,and other information. Table C below shows an example of informationthat may be stored in the deployed monitor database.

TABLE C Monitor ID Location Particle ID Log Images Captured 312945 45Appleseed Drive, 2016-05- 2016-05- Santa Rosa, CA 94555 12_31245_12_31245_ log.txt imageljpg . . . . . . 987431 32 Pear Lane, 2016-05-2016-05- Philadelphia, PA 12_987431_ 12_987431_ 19042 log.txt imagel.jpg. . . . . .

A first column of the table lists an identifier that uniquely identifiesa particle monitor. A second column of the table lists a location wherethe particle monitor is located. In this example, the location includesa street address. The location may instead or additionally includelongitude and latitude coordinates, or any other value or set of valuesthat identifies a geographic location of the particle monitor. A thirdcolumn of the table lists particle identification logs received from theparticle monitor. A fourth column of the table lists particle imagesreceived from the particle monitor.

The particle identification server engine is responsible for performinga server-side analysis of the imaged particles. For example, the cloudserver may have access to computing resources not available locally atthe particle monitor. The particle monitor is designed to be arelatively compact and inexpensive device. The server, however, mayinclude processors more powerful than those at the particle monitor, beable to execute more complex particle identification algorithms than theparticle monitor, and so forth.

In an embodiment, when the particle monitor is unable to identify acaptured particle, the particle monitor notifies the server. The servercan coordinate with the particle monitor in making an identification.For example, the server may use a different set of algorithms to analyzethe particle images transmitted from the particle monitor to the server.Based on the analysis, the server may issue instructions to the particlemonitor for additional images or other data. The instructions mayinclude a request to capture additional images of the particles. Therequest may include a specification of the conditions or parametersunder which the particles should be imaged. For example, the request mayspecify a focal depth at which an image should be taken, illuminationunder which the image should be taken, and so forth.

It should be appreciated that the cloud server is merely representativeof an embodiment. There can be multiple cloud server and storagesystems. Context information or portions of context information may beprovided by one or more third parties. For example, weather conditionsmay be obtained from a third party that offers weather provider services(e.g., AccuWeather).

FIG. 6 shows another block diagram of particle monitor 105 according toone embodiment. In the example shown in FIG. 6, a particle monitor 605includes a housing 610 mounted to a base 615. The housing includes anair intake opening 620, an air exhaust opening 625, and a cartridge slotopening 630. There can be a door connected to the cartridge slot openingvia a hinge. The door can open into the cartridge slot. Shown inside thehousing are a battery or power supply 635 which is connected tocircuitry and logic 640 which in turn is connected to a blower 645,first motor 650, second motor 655, optical subsystem 660, illuminationsubsystem 665, and communications interface 670.

Further shown in FIG. 6 is a removable particle collection cartridge680. The particle collection cartridge includes a reel of tape media.The tape media is wound about the reel and includes an adhesive tocollect airborne particles (e.g., pollen or mold spores). The collectioncartridge is removable from the collection device. That is, a user canremove the cartridge from the collection device without breaking ordestroying the device. There can be an eject button that the user canpress to eject the cartridge from the particle collection device. Forexample, when the collection cartridge is full (or as desired), the usercan remove the collection cartridge from the collection device throughthe cartridge slot opening. The user can then install a new collectioncartridge by inserting the new collection cartridge into the collectiondevice through the cartridge slot opening. The user can then mail theremoved collection cartridge—which contains the collected airborneparticles—to a laboratory for a further in-depth analysis.

The design of the particle monitor and cartridge allows for a veryflexible approach for collecting and analyzing particles. In particular,in another specific embodiment, the cartridge is used for surfaceparticle sampling. Surface particle sampling may be instead of or inaddition to airborne pollen or particle sampling. The cartridgefacilitates a collection system or mechanism that is handheld and easilyportable. A user can hold a body of the cartridge in their hand,position an opening or slot of the cartridge through which a portion ofthe tape is exposed, and press the slot against a surface of an object.Particles on the surface may then be transferred from the surface of theobject to the exposed portion of the tape. The user can then insert thecartridge into the particle monitor for analysis of the particles thathave been collected on the tape.

In a specific embodiment, a handheld portable particle monitor withremovable collection cartridge is provided. In this specific embodiment,the monitor is a relatively small, lightweight, inexpensive, and compactdevice. The monitor is powered by a battery. This allows the monitor tobe easily portable and mobile because the monitor does not have to beconnected to an electrical outlet to operate. A user can take themonitor and cartridge to an environment where there might not be anyelectrical outlets such as to a vineyard, farm, plantation, ranch,forest, or other field environment to collect and analyze airborneparticles, surface-borne particles, or both.

Particles that may be associated with diseases including agriculturaldiseases, plant diseases, animal diseases, and so forth can be easilycollected, analyzed, and identified in the field before widespreaddamage occurs. The handheld particle monitor may include a handleconnected to a body of the monitor so that the monitor can be carried.Instead or additionally, at a least a portion of an outside surface ofthe monitor body may be textured or knurled to facilitate carrying.Further, because the monitor may be used in outdoor environments, aswell as indoor environments, the monitor may include seals to provide aweather-resistance or weather-proof construction. Examples of sealsinclude O-rings, gaskets, all-weather glue, and others.

The particle collection device may include an electronic screen todisplay a status associated with operations of the particle collectiondevice (e.g., “collection cartridge tape 80 percent full,” “analyzingparticles,” “device error,” “transmitting data to remote cloud server,”“firmware update in progress, please wait,” and so forth). There can bestatus lights such as LED status indicators. The particle collectiondevice may include an input device such as a keypad through which theuser can power the device on or off, configure various settings andparameters such as collection frequency (e.g., sample air every 5minutes, every 10 minutes, every 20 minutes, or every 30 minutes), othersettings, and so forth. Instead or additionally, at least some settingsmay be configured remotely such as via a mobile device having a mobileapplication paired with the particle monitor.

The blower may include a fan and is responsible for creating a vacuum inwhich air is sucked into the collection device through the air intakeopening. A flow path of air is directed to the particle collectioncartridge. Particles that may be floating or suspended in the air aretrapped by the adhesive tape of the particle collection cartridge. Theair then exits the collection device through the air exhaust opening.

The first motor operates to rotate the housing of the collection deviceabout the base. The collection device may include an airflow sensor orairflow direction sensing unit that detects a direction of the flow ofthe ambient air. Based on the direction of the airflow, the first motorcan rotate the collection device to orient or align the air intakeopening with a direction of the flow of the ambient air. Instead oradditionally, the first motor may be configured to continuously orperiodically rotate to obtain good representative samples of the ambientair.

The second motor engages the reel of the tape media to unwind theadhesive coated tape media. For example, as airborne particles such aspollen become trapped in a portion of the adhesive coated tape, thesecond motor can unwind the reel to expose a new portion of the adhesivecoated tape upon which new airborne particles can be collected.

The second motor is further responsible for advancing the tapecontaining the trapped particles to the optical and illuminationsubsystems. One or more lighting sources of the illumination subsystemilluminate the trapped particles while a camera sensor of the opticalsystem captures images (e.g., pictures) of the trapped particles foranalysis and identification.

The communications interface is responsible for communications with, forexample, the mobile device, remote cloud server, or both. Thecommunications interface may include an antenna for wirelesscommunication.

FIGS. 7-18 show various views of particle monitor device 105 (andparticle collection cartridge) according to an embodiment. FIGS. 7through 18 illustrate in more mechanical design detail a specificembodiment of an airborne particle monitoring device. FIG. 7 shows anexterior view of a particle monitoring device 700 including acylindrical housing 710 that contains most of the device components aswell as a base 720. Cylindrical housing 710 contains an air-intake slot730 that may be a few centimeters in length and a width that varies fromabout 3 millimeters (mm) to about 1 mm in funnel-like fashion as itpenetrates the thickness of the cylindrical housing 710. The length ofthe air-intake slot may range from about 3 centimeters (cm) to about 10centimeters. This includes, for example, 4, 5, 6, 7, 8, 9, or more than10 centimeters. The length may be less than 3 centimeters.

The cylindrical housing 710 also contains a particle-media-cartridgedoor 740 that may be opened in order to insert or remove particle mediacartridges such as shown in FIG. 8 and discussed below. The air-intakeslot is adjacent or next to the cartridge door. A shape of the cartridgedoor includes a rectangle, perhaps with rounded corners. The cartridgedoor is oriented vertically with respect to a central axis passingthrough the particle collection device. The door may be positionedcloser to the base of the monitor than the top of the monitor.

As shown in the example of FIG. 7, in an embodiment, the cartridge doorincludes a top door edge, a bottom door edge, opposite the top dooredge, and left and right door edges extending between the top and bottomdoor edges. The bottom door edge is closer to the base than the top dooredge and the bottom and top door edges are parallel to each other. Theleft and right door edges are opposite and parallel to each other.

The air-intake slot includes a top intake edge, a bottom intake edge,opposite the top intake edge, and left and right intake edges extendingbetween the top and bottom intake edges. The bottom intake edge iscloser to the base than the top intake edge and the bottom and topintake edges are parallel to each other. The left and right intake edgesare opposite and parallel to each other.

In an embodiment, the air-intake slot is located relatively close to thecartridge door. This helps to allow particles in the air enteringthrough the air-intake slot to be collected on the media cartridge. Forexample, an arc length as measured clockwise (when viewed from above inplan-view) along the outside surface or circumference of the cylindricalhousing from the left door edge to the right intake edge may be A1 (ifviewed from below, it would have been counter-clockwise). An arc lengthas measured clockwise along the outside surface or circumference of thecylindrical housing from the right intake edge to the left door edge maybe A2. Arc A1 may be less than arc A2. A ratio of A1 to A2 may be about1:80. The ratio, however, may vary greatly and in other embodiments maybe about 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:85, 1:90,1:95, 1:100, 1:105, 1:110, 1:115, or 1:120. In an embodiment, theair-intake slot is between a first line or arc about the housingextending from the top door edge and a second line or arc extending fromthe bottom door edge, the first and second lines being parallel to eachother. The air-intake slot may be shaped as a rectangle, oval, obround,circle, or any other shape as desired. There can be multiple air-intakeslots (e.g., two, three, four, five, or more than five air-intakeslots).

The cylindrical housing 710 and its contents may rotate about itscylindrical axis with respect to the base in order to orient theair-intake slot 730 in a desired direction. In some cases, it may bedesired to systematically vary the orientation of the air-intake slot730 in order to average over all directions. Alternatively, the particlecollection device 700 may orient itself so that the air-intake slot 730faces upwind to any breeze or other flow of ambient air. In this lattercase, it is advantageous for the particle collection device 700 toinclude wind or airflow sensors. Visible in FIG. 7 are two of fourwind-detector recesses 750 in which may be mounted airflow sensors insuch a way that they are both exposed to ambient airflow andmechanically protected from accidental impact or contact. In a specificembodiment, a wind-detector recess includes a cantilever deflectiondetector. Wind detectors of many types, including hot-wire airflowdetectors, cantilever deflection detectors, or both may be placed in thewind-detector recesses 750.

The generally cylindrical elongated shape of the housing helps to reduceinterference with other external objects (e.g., grape-vine branches)when the collection device rotates to sample airborne particles such aspollen, mold spores, or both from different directions. In this specificembodiment, a cross-sectional shape of the housing includes a circle. Inother specific embodiments, a cross-sectional shape of the housing mayinclude a square, rectangle, oval, triangle, or any other shape.

FIGS. 8-10 illustrate a particle media cartridge 805 that may be loadedor removed from the particle collection device 700 via theparticle-media-cartridge door 740. The cartridge includes a media forcapturing particles as well as a cartridge body 810. In this specificembodiment the media includes an adhesive coated tape, however, in otherembodiments a different media may be used such as adhesive coatedslides. The cartridge body 810 includes a tape guide structure or wall820 that includes portions including an air-intake zone 830 and aparticle inspection zone 840. The cartridge body 810 includes agear-shaft hole 850 that will be discussed further below. In alternateembodiments, particularly in cases where it is desired to be able torewind as well as advance the tape, there may be a second gear-shafthole (not shown).

FIGS. 9 and 10 show a cross-section of the cartridge body with the media(FIG. 10) and without the media (FIG. 9). The cross-section is for aplane parallel to, in the middle of, planes corresponding to front panel811 (FIG. 8) and back panel 812 (FIG. 8). The dashed lines in FIGS. 9and 10 represent portions of the plan-view edges of front panel 811 andback panel 812 shown in FIG. 8.

Referring now to FIG. 10, a supply reel 1080 of adhesive coated tape1070 is mounted to supply-reel post 960 (FIG. 9). In the air-intake zone830, the tape guide structure 820 fixes the location of the adhesivecoated tape 1070 where it collects particles in the face of air pressurefrom air entering the cylindrical housing 710 (FIG. 7) via theair-intake slot 730 (FIG. 7). The adhesive coated tape 1070 then passesthe particle inspection zone 840 and is finally collected at the uptakereel 1090. Optionally, after use within the particle collection device700, the particle-media cartridge may be removed from device 700 andsent to a laboratory where particles captured by media can be furtherstudied optically or with bio-assays. Such a particle-media cartridgephysically containing captured airborne particles is one embodiment ofphysical archive 260 of FIG. 2.

Referring now to FIG. 8, in a specific embodiment, a user-removable orreplaceable particle media cartridge is provided. The cartridge includesa front panel 811, a back panel 812, opposite the front panel. Sidepanels including a top side panel 813A, a bottom side panel 813B, a leftside panel 813C, and a right side panel 813D extend between the frontand back panels. The top and bottom side panels are opposite to eachother. The left and right side panels are opposite to each other. Thetop and bottom side panels are orthogonal to the right and left sidepanels. As shown for left, top and right sides, side “panels” may coveronly a small portion of their respective side. The cartridge has a shapeof a rectangle.

Referring now to FIG. 10, the right side panel includes a first openingor slot that may be referred to as air intake zone 830. The top sidepanel includes a second opening or slot that may be referred to asparticle inspection zone 840. The left side panel includes a thirdopening or slot that may be referred to as an exhaust port 1079. Alength of the cartridge between the top and bottom side panels is L1, awidth of the cartridge between the left and right side panels is W1, alength of the air intake zone opening is L2, a length of the particleinspection zone opening is L3, and a length of the exhaust opening isL4. In an embodiment, a ratio of L2 to L1 may be about 1:1.5, but mayvary greatly such as 1:1.3, 1:1.4, 1:1.6, or 1:1.7. A ratio of L3 to W1may be about 1:1.4, but may vary greatly such as 1:1.2, 1:1.3, 1:1.5, or1:1.6. A ratio of L4 to L1 may be about 1:1.5, but may vary greatly suchas 1:1.3, 1:1.4, 1:1.6, or 1:1.7.

A thickness of the cartridge between the front and back panels is T1. Awidth of the air intake zone opening is W2. A width of the particleinspection zone opening is W3. A width of the exhaust opening is W4. Inan embodiment, the width of the openings W2, W3, and W4 are equal. Inanother embodiment, a width may be different from another width. In anembodiment, a ratio of at least one of W2, W3, or W4 to T1 is about1:1.4, but may vary greatly such as 1:1.2, 1:1.3, 1:1.5, or 1:1.6. Ashape of the intake zone, particle inspection zone, and exhaust openingsmay be a rectangle or other shape (e.g., oval, round, obround, orcircle).

Inside the cartridge is supply reel 1080, uptake reel 1090, and tapeguide structure 820. The supply reel includes the roll of tape. The tapeincludes an inside or bottom surface 1081A and an outside or top surface1081B, opposite the inside surface. The tape is wound so that the insidesurface faces towards a center of the roll, and the outside surfacefaces away from the center of the roll. The outside surface of the tapeincludes an adhesive. The tape may be made of a thin flexible materialsuch as narrow strip of plastic. In an embodiment, the tape isnon-magnetic or not magnetic or does not include a magnetizable coating.The tape includes an adhesive coating on the outside surface of the tapeto trap particles. In some embodiments, tape may be clear, translucent,transparent, or at least partially transparent to facilitateillumination of trapped particles. That is, the tape may be made of amaterial that allows at least some light to pass through.

The inside surface of the tape may not include the adhesive andpreferably moves with minimal or low friction against tape guide 820.The inside surface may be treated with a coating that allows the insidesurface of the tape to glide freely across the tape guide. For example,in an embodiment there is a roll of tape including an inside surface andan outside surface. A coating or treatment is applied to the insidesurface such that a coefficient of friction of the inside surface afterthe treatment is less than a coefficient of friction of the insidesurface before the treatment. In another specific embodiment, the tapeor portions of the tape may include a magnetizable coating. Such amagnetizable coating may be used to mark and read locations along thelength of the tape of interesting particles that may merit laterlaboratory testing such as bio-assays.

The tape guide structure is sandwiched between the first and secondpanels of the cartridge. The tape guide includes a first segment 1082A,a second segment 1082B, orthogonal to the first segment, and a thirdsegment 1082C extending between ends of the first and second segment.The first segment extends in a direction parallel to the right sidepanel. The first segment extends along at least a portion of the lengthof the front and back panels. The first segment includes a surface thatfaces the first opening (e.g., air intake zone) of the cartridge.

The second segment extends in a direction parallel to the top sidesurface. The second segment extends along at least a portion of thewidth of the front and back panels. The second segment includes asurface that faces the second opening (e.g., particle inspection zone).A length of the first segment may be greater than a length of the secondsegment. A length of the first segment may be less than a length of thesecond segment. A length of the first segment may be the same as alength of the second segment.

The tape extends from the supply reel, across the top surfaces of thefirst, second, and third segments of the tape guide structure, andterminates at the uptake reel. The uptake reel is closer to the top sideof the cartridge than the supply reel. The supply reel is closer to thebottom side of the cartridge than the uptake reel. The tape isconfigured so that the inside surface contacts the top surfaces of thefirst, second, and third segments of the tape guide structure while theoutside surface of the tape, which includes the adhesive, is exposed atthe air intake and particle inspection zones. Thus, particles enteringthe air intake zone can be trapped by the adhesive and then inspected atthe particle inspection zone. The air can pass from the air intake zoneand out the exhaust port of the cartridge. The inside surface of thetape may be smooth or without the adhesive so that the tape can glideacross the tape guide structure.

The first segment of the guide is positioned so that it is slightlyrecessed within the opening of air intake zone 830. That is, right sideedges 1006 of the front and back panels of the cartridge extend slightlypast the first segment. A distance from the right side edges of thepanels to the first segment may be at least a thickness of the tape. Therecessing of the first segment helps to protect the tape from unintendedcontact with other objects.

Similarly, the second segment of the guide is positioned so that it isslightly recessed within the opening of particle inspection zone 840.That is, top side edges 1008 of the front and back panels of thecartridge extend slightly past the second segment. A distance from thetop side edges of the panels to the second segment may be at least athickness of the tape. The recessing of the second segment helps toprotect the tape from unintended contact with other objects.

In the example of the cartridge shown in FIG. 10, the first and secondsegments of the tape guide are on adjacent sides of the cartridge. Thatis the first segment is on the right side of the cartridge and thesecond segment is on the top side of the cartridge. The position of thetape guide segments corresponds to the design of the particle monitor.For example, when the cartridge is inserted into the particle monitor,the second segment of the tape guide will be located directly below theoptical subsystem or microscope including camera sensor and lensassembly. It should be appreciated, however, that the tape guidesegments may be positioned at other locations depending upon the designof the particle monitor.

An angle 1014 is between the second and third segments. An angle 1016 isbetween the first and third segments. In an embodiment, the angles areobtuse, i.e., the angles are more than 90 degrees but less than 180degrees. The angles and positioning of the tape guide segments help toprevent creases in the tape as the tape transitions from the supplyreel, to the intake zone, below and past an upper right corner 1018 ofthe cartridge, to the inspection zone, and to the uptake reel. The endsand corners of the tape guide may be rounded as shown in the figure tohelp ensure that the tape glides smoothly over the tape guide and doesnot snag.

The cartridge, including the tape guide structure, may be made ofplastic, nylon, metal, or other material, or combination of materials.The tape guide structure may be formed or molded as a single unit withone of the front or back panels of the cartridge. Alternatively, thetape guide structure may be formed as a unit separate from the front andback panels. When the tape guide structure is formed as a separate unit,the tape guide structure may be attached to at least one of the front orback panels using any number of a variety of techniques. Such techniquesmay include snap-fits, fasteners (e.g., screws), glues, and others.

Likewise, the front and back panels may be fastened together using anynumber of a variety of techniques. For example, the front and backpanels may be snap-fitted together. The front and back panels may beglued together. In an embodiment, the front and back panels areconnected using screws. In this embodiment, each corner of one of thefront or back panel may include a screw boss. The boss provides amounting structure to receive a screw. The screw passes through a holein a corner of one of the front or back panels and is received by ascrew boss located in a corresponding corner of another of the front orback panels.

FIG. 11 shows a plan-view of selected items of the particle collectiondevice 700 shown in FIG. 7. The device includes two electric motors.Orientation motor 1110 rotates the cylindrical housing 710 and itscontents about its vertical axis and relative to the base 720 (FIG. 7).While the orientation motor 1110 is not centered with respect to theaxis of the cylindrical housing 710, the orientation motor's gear shaft1120 is centered. The intake-reel gear shaft 1140 of the cartridge-reelmotor 1130 extends horizontally and controls the rotation of theuptake-reel 1090 (FIG. 10) of the cartridge. The gear shaft hole 850(FIG. 9) allows the intake-reel gear shaft 1140 to enter theparticle-media cartridge body 810 (FIG. 8). Many of the contentscontained within the cylindrical housing 710, including motors 1110 and1130, are mechanically supported by the internal mounting structure1150. For example, internal components of the monitoring device such aprinted circuit board, motors, and so forth may be attached to theinternal mounting structure using various fasteners, welding, adhesives,or combinations of these. Examples of fasteners include nuts, bolts,screws, and washers. Adhesives include epoxy or glue. Examples ofwelding include plastic welding. The internal mounting structure 1150may be formed of a sculpted volume of plastic. The mounting structuremay be formed using injection molding.

FIG. 12 shows a vertical cross section of particle monitor device 105according to a specific embodiment. In this specific embodiment, theparticle collection device includes three electronic boards. There is amotherboard 1210, an orientation-motor circuit board 1230, and acartridge reel motor circuit board 1240.

Motherboard 1210 contains many electronic components including amicroprocessor (e.g., Raspberry Pi) and a wifi antenna 1220.Alternatively Bluetooth or any other wireless protocol may instead oradditionally be used. For effective wireless communication, it ispreferable that cylindrical housing 710 be constructed from anon-conductive material such as plastic rather than a metal.

Additional circuit boards (not shown) may be included. Also not shown inFIG. 12 for purposes of clarity are numerous wires interconnectingvarious components such as wires between the motors and theircorresponding circuit boards. Motherboard 1210 may contain the hardwareof local processor 240 of FIG. 2.

The motors are located closer to a bottom of the particle monitor than atop of the particle monitor. In an embodiment, orientation motor 1110 islocated near the bottom so as to be close to base 720. Cartridge reelmotor 1130 also is located nearer to the bottom than the top of theparticle monitor as the particle-media cartridge is placed below theoptical system. While the motors may be light weight, in the case thatthe motors are relatively heavy, locating the motors towards the bottomof the particle monitor helps to lower the center of gravity and providestability so that the monitor is unlikely to tip over. Likewise, a powersupply such as a battery may be located closer to the bottom of themonitor than the top of the monitor.

In an embodiment, the motors are light weight. For example, a motor mayweigh about 113-142 grams (or 4-5 ounces) only. In this embodiment, onebenefit of placing the motors on the bottom is because the base of thesystem may be on a surface and when coupled with a ball bearing itenables for easy rotation. The battery is likely to add more weight nearthe bottom which would be advantageous.

Specifically, with respect to a vertical positioning, the orientationmotor is between the bottom of the monitor and the motherboard. Thecartridge reel motor is between the orientation motor and the camerasensor. The camera sensor, being relatively light, is positioned closerto the top of the monitor than the bottom of the monitor. The camerasensor is between the cartridge reel motor and the top of the particlemonitor. With respect to a horizontal positioning, the cartridge reelmotor is between the motherboard and the cartridge door.

FIG. 13 illustrates how sampled ambient air flows through the monitordevice 700. Sampled ambient air 1320 enters through the air-intake slot730 and immediately encounters the air-intake zone 830 (FIG. 10) of theparticle-media cartridge. Here the adhesive-coated tape 1070 (FIG. 10)captures many of the particles within the sampled ambient air 1320.Device-interior air 1330 then exits out the backside of theparticle-media cartridge body 810 (FIG. 8); for this purpose and as seenin FIG. 10, the back side of the cartridge is open rather than closed.Finally exhaust air 1340 (FIG. 13) leaves the device. This airflow isdriven by blower 1310 which pushes out exhaust air 1340 and sucks insampled ambient air 1320. The blower is opposite the air intake slot andabove the exhaust. A gap 1317 between a bottom of the housing and a topof the base allows the exhaust air to escape. The mechanisms for ambientair sampling illustrated in FIG. 13 represents one embodiment of airintake hardware 220 of FIG. 2.

Air intake slot 730 is opposite the blower and is configured to direct aflow path of ambient air created by the blower towards or over the firstopening of the cartridge or air intake zone. For example, there can bechannel, duct, conduit, tube, or passageway that directs the flow pathof the air from the air intake zone. Particles, such as mold spores,pollen, or both, in the air are trapped by the adhesive on the tape.Preferably, the airflow in the air intake zone is turbulent in order tomaximize or increase the chances that particles in the sampled air willbe separated from the air and adhered to the capturing medium. Whendesired, cartridge reel motor 1130 (FIG. 11) advances the tapecontaining the trapped particles to the second opening of the cartridgeor inspection zone. The camera sensor can then capture images of theparticles trapped within the adhesive tape.

FIG. 14A shows a side view of an inside portion of monitor device 700.The monitor device includes an optical subsystem 1405, illuminationsubsystem 1410, cartridge well 1415, platform 1433, and particle-mediacartridge 805. FIG. 14A illustrates a loaded particle-media cartridge inthe cartridge well along with an optical subsystem for particleinspection. FIG. 15 provides more detail on the optical and illuminationsubsystems. During a collection period, particles entering the monitorare trapped within air intake zone 830 by the adhesive of the tape. Thetape or, more specifically, a portion of the tape having the trappedparticles, is then advanced to particle inspection zone 840 forinspection. The duration of the collection period can be configured by auser or administrative user. For example, the collection period may beconfigured to be 5, 10, 15, 20, 30, 60, 90, 120, or more than 120seconds (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 15, 30, 45, or more than 45minutes; 1, 2, 3, 4, 5, or more than 5 hours). The collection period maybe less than 5 seconds.

The optical subsystem includes a camera sensor 1420, lens assembly 1425,and tube 1430. The lens assembly is positioned at a bottom end of thetube and the camera sensor is positioned at a top end of the tube,opposite the bottom end of the tube. The cartridge well receives andholds the particle-media cartridge in a vertical position.

Contributing to the cost-effectiveness of particle-monitoring device 700is the use of a camera sensor 1420 contained within a mass-produced andhighly-integrated camera sensor chip package such as the SONY IMX lineof camera sensors and the Omnivision OV line of camera sensors asprovided by Sony Corporation of Tokyo, Japan and OmniVision TechnologiesInc. of Santa Clara, Calif., respectively. Such highly integratedpackages avoid the cost and mechanical bulk of many associatedelectronic circuits.

As illustrated by the block diagram of FIG. 14B, a highly integratedcamera sensor package 1462 includes within a single chip package notonly the light-sensing pixel sensor array 1464, but also analog driveand readout circuitry 1466, analog-to-digital conversion circuitry 1468,digital image processing circuitry 1470, digital communicationscircuitry 1472 to transfer digital images to a host processor, as wellas many other circuits not shown in FIG. 14B such as power regulationcircuitry, timing circuits, and so forth. With all these supportingcircuits contained with the highly integrated camera sensor package1462, the number of electronic components in the bill of materials for aparticle-monitoring device 700 is significantly reduced, thus reducingcost and enabling a more compact mechanical design.

Platform 1433 is positioned above the cartridge well. The platform canbe between the cartridge well and illumination and optical subsystems.The platform includes a first hole 1435, a second hole 1445, and a thirdhole 1440. The bottom end of the tube of the optical subsystem extendsinto the first hole which opens to face particle inspection zone 840 ofthe particle media-cartridge. In other words, when the particlemedia-cartridge is inserted into the particle monitor, the particleinspection zone of the cartridge aligns with the first hole. The camerasensor is directly above the lens assembly which is directly above theparticle inspection zone. The arrangement allows the camera sensor tocapture images of particles that have been trapped by the adhesivecoated tape.

In other words, in the example shown in FIG. 14A, the platform is abovethe cartridge well that receives the collection cartridge. The camerasensor is positioned within the particle monitor device to be above orover the second opening or particle inspection zone of the cartridge.The camera sensor is closer to a top of the particle monitor than thecartridge.

Positioning the camera sensor above the particle inspection zone helpsto reduce the probability of particles falling onto the camera lens andobscuring the images. For example, in some cases, particles remaining inthe sampled air and not adhering to the tape may settle on the lens, thebond between the adhesive coated tape and collected airborne particlesmay be weak, the adhesive coated tape may include a large collection ormound of particles and particles at the top of the mound may not besecured to the adhesive coated tape, and so forth. The collectioncartridge and camera sensor may be aligned such that a line passingthrough the supply and uptake reels passes through or near the particleinspection zone and lens to the camera sensor. In another embodiment,the tape is transparent and the image capture is from the backside ofthe tape (the non-adhesive side). This prevents particles from enteringthe lens as well since they hit the tape surface and the camera opticsand imaging system is located behind the tape surface.

In a specific embodiment, the cartridge well is rotatable about avertical axis parallel to the central axis passing longitudinallythrough the housing. For example, at least one of the top, bottom, orside of the cartridge well may be connected to a pin (e.g., rod,spindle, shaft, or axle). The pin may sit or revolve within a hole,bushing, or ball bearing connected to the housing. In this specificembodiment, when the media cartridge is loaded through the cartridgedoor and into the monitor, the cartridge well can pivot so that theair-intake slot of the housing aligns with or faces the air intake zoneof the cartridge. This helps to facilitate airflow towards the airintake zone of the cartridge.

In an embodiment, the cartridge well pivots through a distance at leasta thickness of the cartridge. The cartridge well may pivot through anynumber of degrees (e.g., 1, 2, 3, 4, 5, 10, 15, 20, 30, 60, 90, 120, or180 degrees). The ability of the cartridge well to pivot allows theair-intake slot to be located anywhere on the housing. For example, theair-intake slot may be located on an opposite side of the cartridgedoor. In other words, a distance between the air-intake slot and thecartridge door may be equal to a diameter of the housing or half thecircumference of the housing.

Rotating the cartridge away from the cartridge door helps to ensure thatambient or outside light that may enter or leak through the cartridgedoor and into the interior space of the monitor does not enter theparticle inspection zone when the trapped particles are beingilluminated by the illumination subsystem. Reducing or minimizing theamount of ambient or outside light entering the particle inspection zonehelps to ensure accurate measurements.

In a specific embodiment, the illumination and optical subsystems remainstationary or are fixed-in-place while the cartridge well pivots. Thishelps to ensure consistent measurements. In another specific embodiment,one or more of the illumination or optical subsystems may pivot with orwith respect to the cartridge well.

The cartridge well may pivot through any number of positions. Forexample, there can be a first position in which the cartridge well facesthe cartridge door so that the cartridge can be loaded into the well.The cartridge well may then pivot from the first position to a secondposition where the air intake zone of the cartridge faces the air intakeslot of the housing. The cartridge well may remain in the secondposition while the collected particles are illuminated and particleimages captured. The cartridge well can then pivot from the secondposition back to the first position so that the cartridge can be removedand another cartridge inserted.

In another specific embodiment, there can be a first position in whichthe cartridge well faces the cartridge door so that the cartridge can beloaded into the well. The cartridge well may then pivot from the firstposition to a second position where the air intake zone of the cartridgefaces the air intake slot of the housing. Once a collection period hasended, the cartridge well may pivot from the second position to a thirdposition, away from the air intake slot, where the collected particlesare illuminated and particle images captured. The cartridge well canthen pivot from the third position back to the second position foranother particle collection session, or pivot back to the first positionso that the cartridge can be removed and another cartridge inserted.

In other embodiments, the cartridge well may be designed to translate.For example, in another specific embodiment, a particle monitor mayinclude a tray that slides out of the particle monitor. The trayreceives the cartridge and slides back into the particle monitor. In theexample shown in FIG. 7, the cartridge door is shown as being on a sideof the particle monitor between the top and bottom of the monitor. Theside entry helps to facilitate a short overall height and small diameterof the monitor.

In other embodiments, however, the cartridge door may be located on thebottom of the monitor and the cartridge may be inserted through thebottom of the monitor. Locating the cartridge door on the bottom helpsto reduce the probability of unwanted water (e.g., rain) or other debrisentering into the monitor. The cartridge door may be located on the topof the monitor and the cartridge may be inserted through the top of themonitor. Locating the cartridge door at the top can allow a cartridge tobe loaded and removed without having to pick-up the monitor.

As shown in FIG. 14A, the optical subsystem is slightly offset towards aright side of the platform. The optical subsystem is closer to the rightside of the platform than a left side of the platform, opposite theright side. The optical subsystem may be closer to a side of thecylindrical housing than a central axis passing longitudinally throughthe cylindrical housing. This offsetting or arrangement of the opticalsystem helps to facilitate the compact design of the particle monitordevice as other internal components may be located to the left of theoptical tube. Also, to better approximate real-time monitoring, such anoffset also has the benefit of reducing the distance between theparticle capture at air intake zone 830 and particle inspection zone840. In another specific embodiment, the optical axis may be symmetricwith respect to the light sources.

The second hole 1445 houses a first illumination or light source 1450.Light from a first light emitting element 1524 (FIG. 15) is directedwithin a guide 1526 to a diffuser 1525. The third hole 1440 houses asecond illumination or light source 1455. The illumination sourcesilluminate the particle inspection zone so that the camera sensor cancapture images of the trapped particles when illuminated by theillumination sources.

For example, FIG. 15 shows an enlarged view of a portion of the sectionview shown in FIG. 14A. Particles 1530 collected on the tape have beenmoved to particle inspection zone 840. First light source 1450illuminates 1503 the particle inspection zone with first light. Camerasensor 1420 (FIG. 14A) captures images of particles in particleinspection zone 840 (or a scene) within a field view of the camerasensor to generate a first image. That is, the first image is generatedwhile the particles are illuminated with the first light. Second lightsource 1455 illuminates the particle inspection zone with second light,different from the first light. The camera sensor captures particles inthe particle inspection zone within the field view of the camera sensorto generate a second image. That is, the second image is generated whilethe particles are illuminated with the second light. The first lightsource may be deactivated when capturing the second image.

In an embodiment, it is desirable that the area around the particleinspection zone be dark. This helps to provide a controlled lightingenvironment for illuminating the particles under different specifiedlighting conditions and image capture. Thus, components within the areaaround the particle inspection zone may be black, colored black (e.g.,painted black or a dark color), non-reflective, processed so that theresulting surface finish is darker as compared to the surface before theprocessing, and so forth.

The first and second lights have different spectral characteristics. Forexample, the first light may include white light (e.g., light having abroad range of wavelengths and is perceived by the human eye as beingcolorless) and the second light may include light corresponding to anabsorption spectrum of a particle of interest, or the second light mayprovide ultraviolet light illumination capable of exciting fluorescenceof biomolecules. In an embodiment, the first and second images areanalyzed to identify or discriminate the particles. For example, thefirst and second images may be compared to each other to detect changesor differences in the appearance of the particles in the images based onthe different lighting conditions under which the particles werephotographed.

Detecting such changes (or the lack of changes) can provide anindication of what a particle might be (or not be) because differenttypes of particles can have different light absorption characteristics.These differences in light absorption characteristics can be exploitedin order to identify or discriminate the particles. Capturing variousimages of the same particles but under various different lightingconditions can be used to “probe” and identify or discriminate theparticles.

As discussed, lens assembly 1425 images the particles within particleinspection zone 840 on camera sensor 1420 (FIG. 14A). The lens assemblymay include one or more lenses. FIG. 15 illustrates the case where thelens assembly includes a weak (i.e., longer focal length) lens 1512 incombination with a strong (i.e., shorter focal length) lens 1514. In theexample shown in FIG. 15, the strong lens is closer to the particleinspection zone than the weak lens. Optionally, the lens assembly 1425may provide an electrically controlled focal length, for example,through a combination of a strong lens 1514 of fixed focal length and aweak lens 1512 whose focal length is electrically controlled.

Given a fixed location of camera sensor 1420 and of lens assembly 1425,increasing the net or effective focal length of the lens assembly 1425moves the object focal plane down and decreasing the focal length movesthe object focal plane up. That is by properly adjusting the focallength of the lens array 1425, one can bring into focus particles 1530.Furthermore, for larger particles or for optical arrangements withshallower depths of field, different adjustments of net focal length ofthe lens assembly 1425 can bring into focus different horizontal layersof a translucent particle like pollen grains. A set of images focused ondifferent horizontal layers may provide information on thethree-dimensional structure.

A lens with an electrically controlled focal length is generally morereliable than a moving mechanical mechanism. In other words, thereliability of modern electronic devices depends heavily in replacingmoving mechanical mechanisms with electronic mechanisms. From thisperspective, it is very attractive to be able to be able to adjust inreal-time the focus of the particle monitor's optical system with no orfew mechanical movements, but instead control the focal length of thelens assembly purely electronically. Available lenses withelectronically controller focusing tend to be weak lenses, too weak foridentifying or discriminating particles. A weak lens in combination witha strong lens, however, can provide for reliability identifying ordiscriminating particles. In other words, this problem can be overcomewith a strongly focusing lens assembly comprising a fixed strong lensand a weak electronically controlled lens.

In any case, an optical axis 1515 of the lens array intersects theparticle inspection zone 840 where particles 1530 such as mold spores,pollen grains, or both may be located. Such a lens assembly may be partof image capture hardware 226 of FIG. 2.

The camera sensor 1420 (FIG. 14A) may be a black-and-white camerasensor, but in order to generate richer spectral information it ispreferable that the camera sensor 1420 be color sensitive such asproviding the ability to capture RGB (red-green-blue) color images.Indeed, the large volume or scale at which color camera sensors aremanufactured as compared to black-and-white camera sensors have resultedin color camera sensors being less expensive than black-and-white camerasensors.

Both black-and-white cameras and color cameras provide information onthe shape and structure of imaged objects, in other words the“morphology” of imaged objects. Color cameras also provide colorinformation. The particle monitor can analyze an image to distinguishbetween types of particles through morphological features (e.g., is itround or rod like?, is it smooth or spikey?, is it large or small?, andso forth).

In another specific embodiment, the camera sensor 1420 may be alight-field camera sensor. These items represent embodiments of theimage capture hardware 226 of FIG. 2.

Referring back now to FIG. 15, third hole 1440 within platform 1433houses second illumination source 1455. The second illumination sourcemay include, as shown in FIG. 15, a second light emitting element 1540,a quantum-dot film 1555, an optional diffuser 1550, and an optical shaft1560. Second hole 1445 within the platform houses first illuminationsource 1450. A diffuser can be optional as the quantum-dots themselveswill randomize the directions of emitted light. Alternatively,quantum-dot film 1555 may be absent and second light emitting element1540 may directly illuminate the particle inspection zone 840.

Second light emitting element 1540 provides light reaching the particleinspection zone 840 via light propagation approximately parallel to anillumination axis 1545. The illumination light may be visible light, UVlight, or infrared light, or a combination thereof. As discussed,different types of particles can have different light absorptioncharacteristics. For morphology analysis, visible light, or even onecolor of visible light can be sufficient. It is also an option toperform morphology analysis based on UV fluorescence images. In otherwords, UV fluorescence images may be used for morphology analysis. Thus,a UV light source may be used to identify a shape and outline of theparticle and to also probe its fluorescent properties orcharacteristics. As a result, in an embodiment a visible or white lightsource may be omitted from a particle monitor.

In some cases, a morphology analysis will not be sufficient to make aconclusive identification as there can be particles of different typesbut which have the same or similar geometric features. Color informationbecomes particularly interesting when it provides even a crude level ofbiochemical analysis without the delays and cost of wet-laboratorytechniques. The differences in light absorption characteristics ofdifferent particles can be exploited to identify particles ordiscriminate between particles.

For example, pollen grains tend to have a yellowish color, so color asperceived by the human eye, or an RGB camera sensor under white lightillumination is of value to check if a candidate pollen grain is indeedyellowish. Illuminating with white light and capturing the resultingimage provides a useful indication of the colors of the particles thathave been captured. Grass pollens tend to have bio-moleculechlorophyll-a and hence a pollen grain with visible light absorptionpeaks of chlorophyll-a is likely to be a grass pollen.

Fluorescence under UV illumination is a marker of bio-molecules that canbe used to distinguish between organic and inorganic particles.Biochemical information can be provided by UV fluorescence. Fluorescenceis a property some molecules have in which they absorb light of onecolor and emit light of a different color (e.g., different wavelength).While UV light might not be detected by the camera sensor, the resultingfluoresced or emitted light from the particle may be detected by thecamera sensor. As another example, illumination in near infra-red (nearenough in wavelength to visible light to be detected by the camerasensor) may provide useful information in regards to identifyingparticles or discriminating between particles.

Camera sensor 1420 (FIG. 14A) may image scattered light, fluorescentlight, or both. Light scattering, where photons bounce off objects in adifferent direction without changing their wavelength/color is theworkhorse of light imaging. With rare exceptions, this is how we seeobjects in our daily lives when we use our eyes. Just as with our humaneyes, in the particle monitor's basic object shape and color informationcomes from light scattering. UV fluorescence is less common (such as in“black lights”) and is of interest in particle identification anddiscrimination because UV fluorescence indicates the presence ofbiological materials and may provide information about the types ofbiological materials.

Optionally, to provide a more uniform illumination of the particleinspection zone 840, a diffuser 1550 may be placed along theillumination axis between second light emitting element 1540 and theparticle inspection zone 840. The second light emitting element 1540 andthe diffuser 1550 may be mechanically connected with an optical shaft1560 forming a rigid illumination-source sub-assembly.

Preferably, optical shaft 1560 has optical wave-guiding properties so asto more efficiently direct light from second light emitting element 1540to particle inspection zone 840. Third hole or illumination channel 1440may penetrate platform 1433 in order to hold the rigidillumination-source sub-assembly in place and to remove material aroundthe illumination axis 1545.

As shown in the example of FIG. 15, there is a 55-degree angle betweenthe optical axis 1515 and the illumination axis 1545; however otherangles are also possible. This includes angles larger than 90-degrees ifthe adhesive-coated tape and the tape guide 820 under the particleinspection zone 840 are at least partially transparent. Larger than90-degree illumination angles are also an option for alternateembodiments for which sampled particles are captured on transparentglass or plastic slides instead of with adhesive-coated tape. FIG. 15illustrates just one possible azimuthal angle for an illumination axis1545; optionally any azimuthal angle may be used. Such illuminationoptions represent embodiments of illumination hardware 224 of FIG. 2.

Light emitting element 1540 may be an LED (light emitting diode)including possibility an OLED (organic light emitting diode), or alaser, or any other type of light generating device. Furthermore, lightemitting element 1540 may be of the downstream end of an optical fiberbringing light from a light emitting element mounted elsewhere. Lightemitting element 1540 may provide a wide range of wavelengths, such aswith a white-LED, or provide a narrow range of wavelengths, such as witha laser. To provide more information for recognition of particle types,there can be multiple illumination sources.

The holes formed in the platform for light sources, optical shaft, orboth may have a cross-sectional shape of a circle. In other embodiments,the cross-sectional shape of a hole may be an oval, square, rectangle,or other shape. In some applications it may be useful to usecross-sectional hole shape as part of a keying system that controls whattype of illumination source sub-assemblies are inserted into whichholes. A light source may include a light emitting element and opticalfiber. The use of optical fiber allows the light emitting element to belocated anywhere within the particle monitor and not necessarily withinthe platform. The ability to locate the light emitting element anywherewithin the particle monitor helps to facilitate a compact design.

For example, the light emitting element may be located in the base ofthe platform. An end of an optical fiber may be connected to the lightemitting element. An opposite end of the optical fiber may be connectedto a hole or opening in the platform. The optical fiber transmits lightfrom the light emitting element to the platform or particle inspectionzone so that the collected particles can be illuminated for the camerasensor. There can be multiple strands of optical fiber. Across-sectional shape of the optical fiber may be a circle or othershape.

As previously stated, image capture hardware 226 of FIG. 2 mayadvantageously take advantage of commercially available camera sensors.Due to mass market demand for color digital cameras including camerasbuilt into smart phones, sophisticated RGB (red, green, blue) camerasensors are available at relatively low cost. A feature of the systemallows for the use of such relatively low-cost camera sensors for anautomated pollen detection systems based on optical imaging of sampledpollen. Spore and pollen color information is of interest fordifferentiation between types of biological particles. Now let us take acloser look at the spectral characteristics mass produced camerasensors.

FIG. 16 illustrates spectral characteristics typical of an RGB camerasensor such as may be found in consumer-level digital cameras.Horizontal axis 1610 represents optical wavelengths within the visiblespectrum from violet at the left to red at the right. Vertical axis 1620represents the quantum efficiency (percentage of photons resulting inone electron of current) of a camera sensor sub-pixel. Thedot-dot-dashed curve 1630 represents the spectral response of red or “R”sub-pixels of an RGB camera sensor. The dot-dashed curve 1640 representsthe spectral response of green or “G” sub-pixels of an RGB camerasensor. The dashed curve 1650 represents the represents the spectralresponse of blue or “B” sub-pixels of an RGB camera sensor. Note thatthe quantum efficiency goes to zero for wavelengths shorter than about380 nm, or stated more simply, RGB camera sensors generally to notrespond to ultraviolet light.

These spectral responses are determined in large part by color filtersplaced in front of sub-pixels as part of the construction of the camerasensor. Often, the color filters include more green elements as comparedto red or blue elements. This is because the human visual system peaksin sensitivity in the green spectral region (e.g., peaks atapproximately 550 nm wavelength). Thus, the abundance of green sensorpixels in the imaging device allows for approximating the color responseof the human visual system.

As can be seen, the spectral response curves are quite broad andoverlapping. For conventional digital camera purposes, this has theadvantage that there is no visible light wavelength for which a colordigital camera is blind. However, from the perspective of quantitativespectral analysis, the broad and overlapping spectral characteristics isa disadvantage because the absorption characteristics of particles ofinterest (e.g., grass pollen) may be much more narrow. Thus, in somecases, it can be very difficult to distinguish and discriminatedifferent particle types based on color using a broad and overlappingemission spectra to illuminate the particles.

A close look at the spectral profiles in FIG. 16 reveals a significantamount of what may be described as “color crosstalk.” For example, evenred light at a wavelength of 700 nm will not only excite red RGB pixelswith a quantum efficiency of about 35 percent, but also excite “green”RBG pixels with a quantum efficiency of order 10 percent and excite“blue” RGB pixels with a quantum efficiency of order 5 percent. It is anover-simplification to say that “red” RGB pixels detect “red” light,“green” RGB pixels detect “green” light and “blue” RGB pixels detect“blue” light. For a deeper appreciation of the system discussed herein,and in particular various embodiments of the illumination hardware 224(FIG. 2) presented further below, it is useful to keep this fact inmind.

In conventional applications of RGB camera sensors, such as in colordigital cameras, color digital microscopes, and so forth, it is takenfor granted that associated lens assemblies must be achromatic so thatthe red, green and blue sub-pixel images are all brought to an equallysharp focus. RGB camera sensors are conventionally associated withachromatic optics. The requirement that RGB camera optics be achromaticadds to the complexity of the optics, and hence its cost, particularlyif a relatively large aperture is required.

Fortuitously, the spectral widths that are possible from illuminationsources using quantum dots is a good match to the spectral widths of,for example, the absorption peaks of chlorophyll-a as shown in FIG. 17.FIG. 17 shows an absorption spectrum of chlorophyll-a. With a verticalaxis 1710 representing absorption strength and a horizontal axis 1720for light wavelength, a curve 1730 shown in FIG. 17 represents theabsorption spectra of chlorophyll-a.

This absorption spectrum has a pronounced “chlorophyll-a red peak” 1740and a pronounced “chlorophyll-a blue peak” at 1750. In the plot of FIG.17, the two absorption peaks are centered near 665 nm and 465 nmwavelengths. As one of skill in the art would recognize, the solventsolution environment of the chlorophyll-a has a significant effect onthe locations of the peaks. Thus, depending upon factors such as thesolvent solution environment, the location of the peaks may differ suchas for chlorophyll-a on grass pollen. It should be appreciated that theprinciples of choosing quantum-dots based on the locations of the peaksremain the same.

The presence of chlorophyll-a distinguishes grass pollens from otherpollens as well as other particles, and hence a quantum-dot illuminationsource tuned to an absorption peak of chlorophyll-a is of interest inthe identification of allergenic grass pollens. Like plants, fungi alsoproduce pigments with distinctive colors, such as the bright red ofdeadly red cap mushrooms. In an embodiment, quantum-dot illuminationsources may be tuned to characteristic absorption peaks of certainfungal spores.

FIG. 18 shows a top view of a platform 1805 of another specificembodiment of a particle monitor. FIG. 18 illustrates an example witheight illumination sources or channels 1871, 1872, 1873, 1874, 1875,1876, 1877, and 1878. The illumination channels have been drawn in FIG.18 with varying shapes to represent different azimuthal and polarangles. In FIG. 18, variations in polar angle result in variations indistances from center in plan view. For example, illumination sources1872 and 1875 have the same polar angle but 180-degree oppositeazimuthal angles.

Each illumination channel may include a light emitting element and anoptical shaft. For example, illumination channel 1877 includes a lightemitting element 1885 connected to an optical shaft 1886. Illuminationchannel 1878 includes a light emitting element 1887 connected to anoptical shaft 1888. An illumination source may or may not be associatedwith a set of quantum dots. An illumination source may emit visiblelight (e.g., wavelengths ranging from about 390 nm to about 700 nm), UVlight (e.g., wavelengths ranging from about 10 nm to about 380 nm), orinfrared light (e.g., wavelengths ranging from about 700 nm to about 1mm).

The light emitting elements and optical shafts for the remainingillumination channels have been omitted for clarity. In other words,additional light emitting elements (not shown) may be installed inadditional illumination channels 1871-1876. Each illumination source hasan illumination axis that intersects the particle inspection zone 1840.Illumination axes corresponding to different illumination sources mayvary in azimuthal angle as well as angle with respect to an optical axispassing through a particle inspection zone 1840 having particles 1830,through a lens assembly, and to a camera sensor. Having many differentillumination axes further provides for additional dimensions ofanalysis. For example, the lengths of different shadows resulting fromshining light at different angles can indicate the height of a particle.

Compared to fluorescence images under UV light illumination, images fromvisible light scattering are more sensitive to the illuminationdirection. This is because while incoming directions of UV excitationphotons have little influence on the outgoing directions offluorescently emitted photons, the incoming directions of illuminatingvisible light photons have a strong influence on the outgoing directionof corresponding scattered photons. For example, placement of a UV lightsource in illumination channel 1877 results in essentially the samecamera sensor fluorescent-light image as for the placement of the UVlight source in illumination channel 1878.

However, placement of a white visible light source in illuminationchannel 1877 results in a strikingly different camera sensor image ofvisible scattered light as for the placement of the white visible lightsource in illumination channel 1878.

An everyday example of this effect is that waves on a lake sparkle verydifferently when the sun is near the horizon than at high noon. Thisanalogy with light scattering off water surfaces is not far afieldbecause fungal spores and other biological particles of interest areoften transparent like water or semi-transparent under visible light.Visible light will often be scattered at the air/spore-material boundarysurface much as light is scattered from air/water boundary surfaces. Thenature of visible light scattered from a transparent spore is similar tothe nature of light scattered from a transparent glass object.

Depending on lighting conditions, some surfaces of glass objectionsdon't reflect light towards our eyes is not visible to us. Similarly,reflected light images of spores may give a complete picture of sporegeometry. Camera sensor images of scattered visible light are stronglybiased by the illumination direction of the light source, whilefluorescent light images are largely independent of the illuminationdirection of the UV light source. The illumination direction bias of thevisible light image complicates comparisons of fluorescent-light imageswith scattered-visible-light images.

Using multiple illumination directions may significantly reduce theillumination-direction bias of scattered-visible-light images of camerasensor 1420. For example, in one embodiment, a UV light source is placedin illumination channel 1878 while three separate white-light LEDs areplaced in illumination channels 1872, 1875 and 1877. By way of example,directions of illumination channels 1875, 1877, and 1872 may beseparated from each other by right angles in the plan view of FIG. 18and vary in polar angles; other directions and other numbers ofwhite-light LEDs may also be used. Also visible-light sources that arenot white-light LEDs may also be used, such as a red laser (resulting invisible light scattering data being largely contained in red RBG camerasensor pixels while green and blue fluorescence signals are largelycontained in blue and green camera sensor pixels). By combiningscattered light signals corresponding to the three visible-lightillumination directions, the illumination-direction bias is muchreduced. Several means are available to combine the scattered lightsignals. White light sources in illumination channels 1872, 1875 and1877 may be turned on simultaneously during image capture.

Alternatively, in another specific embodiment, the three white-lightsources are turned on sequentially during the camera sensor exposureperiod. In this specific embodiment, a particle monitor includes aparticle inspection zone and first, second, and third white-lightsources above the particle inspection zone. The first, second, and thirdwhite-light sources may be arranged to provide illumination of theparticle inspection zone at different angles. A first distance is fromthe particle inspection zone to the first light source. A seconddistance is from the particle inspection zone to the second lightsource. A third distance is from the particle inspection zone to thethird light source. The first distance may be different from the seconddistance, third distance, or both. The first distance may be the same asthe second distance, third distance, or both.

The first, second, and third light sources may form first, second, andthird vertices of a triangle. A first side of the triangle is betweenthe first and second vertices. A second side of the triangle is betweenthe second and third vertices. A third side of the triangle is betweenthe third and first vertices. Lengths of the first, second, and thirdsides of the triangle may be equal (e.g., equilateral triangle). Alength of at least one of the first, second, or third sides may be thesame as or different from a length of at least another of the first,second, or third sides.

A method may include activating the first light source and capturing afirst image while the first light source is activated; activating asecond light source and capturing a second image while the second lightsource is activated; and activating a third light source and capturing athird image while the third light source is activated. During thecapturing of an image, two or more light sources may remain activated.For example, during the capturing of the second image, the first andsecond light sources may remain activated. Alternatively, during thecapturing of the second image, the first light source may be deactivatedwhile the second light source remains activated. During the capturing ofthe third image, the first, second, and third light sources may remainactivated. Alternatively, during the capturing of the third image, atleast one of the first or second light sources may remain deactivated.

A further option is to capture and separately digitize the visible lightimages corresponding to the three different illumination directions andthen digitally combine the three images in software. By whatever means,combing signals for multiple white-light illumination directionsprovides a less biased scattered visible light image for use indetermining outlines of particles and for comparing with UV fluorescentimages.

The light emitting elements may vary in the nature of their emittedlight. For example, illumination hardware 224 (FIG. 2) may provide localprocessor 240 many illumination options such as white-lightillumination, UV illumination, infrared illumination, and visible lightilluminations of various color characteristics. Local processor 240 mayactivate individual light sources one-at-a-time, or activate two or morelight emitting elements simultaneously or concurrently.

FIG. 19 shows a plot combining camera-sensor sub-pixel spectralcharacteristics as shown in FIG. 16 with illumination source spectralcharacteristics. Vertical axis 1907 represents the quantum efficiency(percentage of photons resulting in one electron of current) of a camerasensor sub-pixel. Horizontal axis 1909 represents optical wavelengthsspanning from the near ultraviolet to the left, the visible spectrumfrom violet at the left to red in the middle, and near infra-red to theright. The horizontal axis of FIG. 19 is expanded relative to thehorizontal axis of FIG. 16. This was done to include UV and infra-redlight sources.

The dot-dot-dashed curve 1930 represents the spectral response of red or“R” sub-pixels of an RGB camera sensor. The dot-dashed curve 1940represents the spectral response of green or “G” sub-pixels of an RGBcamera sensor. The dashed curve 1950 represents the represents thespectral response of blue or “B” sub-pixels of an RGB camera sensor.FIG. 19 shows the quantum efficiency curves for red, green and bluecamera sensor sub-pixels. Relative to FIG. 16, the horizontal wavelengthaxis has been extended in FIG. 19 to include ultraviolet (UV) light atshorter wavelength and near infrared (IR) light at longer wavelengths.

FIG. 19 adds emission spectra of value illumination sources (for whichthe vertical axis has arbitrary units unrelated to quantum efficiency).Heavy solid curve 1910 is representative of the emission spectra ofwhite-light LEDs. White-light LEDs are fundamentally blue LEDS, hencethe emission peak in the blue, to which phosphors have been added toconvert much of the blue light into a broad spectrum of longerwavelength light, hence the broad spectral peak to the right.

In many embodiments, such a white-light LED is the primary illuminationsource used to produce images for particle shape (morphology) analysisas well as a basic, first pass color analysis. This first pass coloranalysis is largely based on color as perceived by the human eye. It isworth keeping in mind, there is much more color information that can beperceived by the human eye.

Curve 1920 represents the emission spectra of a blue LED. While awhite-light LED may be used to excite fluorescence of quantum-dots, itis more efficient to do so with a blue LED.

Curve 1932 represents the emission spectrum of quantum dots tuned duringmanufacture to emit red light at the absorption peak of chlorophyll-awithin grains of grass-pollen. This “chlorophyll-a red” emission may befluorescently exited by, for example, light from a blue LED, or exciteddirectly electronically. Curves 1934 and 1936 illustrate spectra ofquantum-dots tuned to emit light of wavelengths just above and justbelow the chlorophyll-a red wavelength.

A strong signature for the presence of chlorophyll-a is strong opticalabsorption of red light of the spectrum of curve 1932 but not of redlight of the spectrum of curves 1934 and 1936.

An approximation of full spectral analysis of objects viewed with acamera sensor is possible with a sufficient number of quantum-dotillumination sources. Consider, as an example, extending the set ofspectral curves 1934, 1932 and 1936 in both directions of increasing anddecreasing wavelength in order to cover the entire visible spectrum.While not providing the same fine color resolution of a scientific gradespectrometer, a device with between 10 and 100 quantum-dot illuminationsources may still provide an approximation of a full spectral analysisat each camera-sensor pixel location that provides useful information atrelatively low cost.

Even for analysis of shape information (morphological analysis) thatdoes not make use of color, the narrow spectral widths of quantum-dotemission may be helpful. Consider, as an example, a lens system that issubject to chromatic aberration, either as a cost saving measure or dueto the use of an electronically controlled variable lens (in combinationwith a stronger fixed lens). In such a scenario, illumination with thegreen quantum-dot spectrum of curve 1945 will largely eliminatechromatic aberration effects and produce sharper images formorphological analysis.

Useful spectral information is not limited to the visible spectrum. Forexample, it may be of interest to illuminate particles of interest witha near infrared LED, for example, at a wavelength of 850 nm. Asillustrated by curve 1955, sufficiently “near” infrared light, that iswith sufficiently short wavelengths, may still be transmitted by commonlens materials and be detectable by a conventional camera-sensors. Insome applications, the near infrared properties of particles may be ofvalue.

Typically, common lens materials block ultraviolet light. This may beused to advantage when particles of interest are illuminated by UVlight, resulting in fluorescent light of longer wavelengths that aredetected by the camera-sensor while the illuminating UV is not. Thisisolates the interesting fluorescence signal from simply scattered UVlight. UV fluorescence is of particular value in distinguishing betweeninorganic particles and particles of biological origin.

Curve 1960 is representative of common 365 nm UV LEDs. This UVwavelength is sufficiently short to fluorescently excite flavins such asriboflavin and related biomolecules, but too long to strongly excitemost other bio-molecules, hence a 365 nm UV LED can be used to probeflavin content of biological particles of interest.

Curve 1962 represents the emission spectra of a shorter wavelength UVLED, with the ability to fluorescently excite trytophan, an aromaticamino acid within proteins. Even shorter UV wavelengths may be used tofluorescently probe further sets of biomolecules.

By probing both flavins and protein content, a pair of UV LEDs providesa two-dimensional probe of particle biochemistry. In a like manner andfor similar purposes, additional UV LEDs may be included where each UVLED (or cluster of UV LEDs) emits UV light of differing energies. Whileless powerful than a wet-laboratory bio-assay, such optical probing ofparticle biochemistry has the advantage of providing immediate, ifcrude, biochemical information, with which to aid in real-time particletype discrimination. Because biomolecules have varying degrees ofstability, particularly those with conjugated bonds associated withfluorescence, their presence or absence may correlate with the metabolichealth of a biological particle such as a fungal spore. Thus, UVfluorescence images can be analyzed by the particle monitor to not onlymake an identification of the species of a biological particle such as afungal spore, but to also distinguish between different states of healthof the biological particle.

FIG. 20 shows a flow chart illustrating some basic ingredients ofautomated particle (e.g., spore) monitoring according to a specificembodiment. This flow chart illustrates a method of spore detection. Inbrief, in a step 2010, a spore monitor samples ambient air such as via ablower that sucks in the ambient air. In a step 2020, mold spores,pollen, or both captured from the ambient air is transported to anillumination zone. For example, there can be a supply reel and a take-upreel to transport a sticky tape with adhesive side up past theair-inlet/blower. Fungal spore and other particles that stick to theadhesive are transported to the illumination zone which is provided byone or more illumination sources, some of which may be ultravioletillumination sources.

In a step 2030, one or more illumination colors are selected. In a step2040, the pollen is illuminated. The selection of the illuminationcolors may be based on a pre-determined illumination sequence that isstored by the monitor. The particle (e.g., spore) monitor may access thepre-determined illumination sequence in order to identify the color(e.g., wavelength) of light that should be emitted. The selection may becontrolled by a computer. The computer selects an illumination source(s)with desired spectral properties. The selected illumination sources arethen activated.

In an embodiment, the illumination sequence may be determineddynamically. A method may include illuminating the captured particlesunder white light, while the particles are being illuminated by thewhite light, capturing a first image of the particles, identifying, fromthe first image, colors of the particles, based on the colors of theparticles as revealed by the first image, and selecting another color,different from white, with which to illuminate the particles for asecond image of the particles. For example, RGB images collected underwhite light illumination may contain yellow particles of a shapepossibly indicative of grass pollen grains. As discussed previously, theinterpretation of the imaged pollen grains being grass pollen may thenbe tested by using quantum-dot illumination sources corresponding tospectral curves 1932, 1934 and 1936 of FIG. 19.

In a step 2050, the monitor performs an optical detection. The opticaldetection may include capturing an image of the particles under theillumination. In other words, while the sampled spore or pollen isilluminated, a lens array and an RGB camera sensor capture images of thesampled spore or pollen.

Steps 2030-2050 may be repeated 2052 any number of times in order tocapture further color information about the sampled particles (e.g.,pollen or spores) and other detected particulates. In a step 2060, theoptical data (e.g., images of particles) is analyzed. In a step 2070,the optical data (e.g., images) may be transmitted to a remote server.

In an embodiment, as discussed above, the particle monitor can beconnected to a network. The connection to a network allows the particlemonitor to receive updates. An update may include, for example, updatesto the illumination sequence, updates to image capture settings, orboth. An illumination sequence stored at the particle monitor mayspecify the order for activating the different illumination sources. Theimage capture settings may specify focal depths or depths of focus. Forexample, three-dimensional morphology information may be obtainedthrough a sequence of depths of focus corresponding to differenthorizontal layers of a translucent particle such as a fungal spore. Theability to update the particle monitor remotely or over a network helpsto ensure use of the latest algorithms for quickly and accuratelyidentifying or discriminating particles.

In a specific embodiment, aspects and principles of the system may beapplied to monitoring a vineyard for agriculture diseases oragricultural pathogens. In this specific embodiment, the collectioncartridge can be hand-held by a user for a manual collection ofparticles that may have collected on a surface of a leaf or grape of agrape vine. In this specific embodiment, a method may include holding acollection cartridge, the collection cartridge comprising an adhesivecoated tape and a slot through which a portion of the tape is exposed;positioning the slot to face an object; pressing the cartridge againstthe object to bring the portion of the tape into contact with a surfaceof the object, thereby transferring particles on the surface to thetape; and inserting the cartridge into a particle monitor for ananalysis of the particles. The object may include a leaf, such as agrape leaf, or a grape such as from a grape vine. The types of particlesof interest to identify may include small pests, insects, bacterium,mildew, fungal spores including mold spores, or combinations of these.

Examples of airborne fungal spores of interest to vineyards may includepowdery mildews such as Erysiphe necator, Eutypa Lata, Botrytis, andCladospora mold among others. Mildew is a fungus. Powdery Mildew(Erysiphe necator) is widely considered to be the most problematic ofall the vineyard molds. In an embodiment, the particle monitor isconfigured to discriminate the spore health of powdery mildew.

Detection of such mold may be transmitted to a mobile app on thevineyard owner's mobile device. The system can provide counts, trends,and predictive data and analytics displayed via a web application ormobile application. The application allows for customizing alerts forefficient vineyard management operation. The system can provideup-to-the minute information on invasive, disease causing molds,pollens, and weeds. Winds, for example, can carry disease spores formiles. It is desirable to distinguish between harmful and benign moldsfor successful fungicide operations. The system allows for 24/7monitoring and is much more cost-effective than microscopic inspectionand visual spot checks. Early disease detection and control can increaseyield and product quality.

In an embodiment, systems and techniques are provided for the detectionand classification of airborne particles (e.g., pollens, molds, dander,heavy smoke (ash) particles, sand/silica, asbestos, and many others).Systems and techniques are provided for detecting and counting particleshaving a size (e.g., a longest dimension) from about 1 um to about 1500um. In an embodiment, a minimum particle resolution is about 0.3 um. Inanother embodiment, a minimum particle resolution is about 0.1 um). Inan embodiment, a light-based methodology includes multiple differentanalysis techniques including deep neural network machine learning andadvanced algorithms to extract unique particle signatures leading toclassification.

A media cartridge is provided that captures particles for physicalrecord archiving, future studies, advanced studies in a laboratory, orcombinations of these. An analysis may include particle featureextraction, vector extraction, executing a classifier algorithm,particle classifications, and aggregating the information into a resultsfile, or combinations of these. The results file may be transmitted to auser's mobile device for display. Particle detection techniques mayinclude morphology (e.g., shape and size), UV fluorescence (e.g.,flavin, NADH & protein excitation), colorimetry (e.g., colorparameters), topography (e.g., height and texture), internal structure,or combinations of these.

Particle monitoring apparatus and methods have been described above thatprovide systems and techniques to identify the types of airbornebiological particles. For example, airborne particles may be identifiedas a spore of a particular species of fungus or a pollen grain of aparticular type of flowering plant. Advantageously, such apparatus andmethods may be used to not only identify the types of biologicalparticles, but also recognize the state of biological particles. Forexample, in the control of agricultural pathogens, when a particulardisease causing fungal spore is detected, it is of interest to know ifthe spore is in a healthy and hence virulent state, or in a sterilestate as may result from exposure to a fungicide.

FIG. 21A shows a flow chart describing the extension of particleidentification to the determination of the state of a particle. At step2103, the type of a biological particle has been determined withapparatus and methods such as described above. For example, the particlemay be identified as a fungal spore of powdery mildew or as a pollengrain of ragweed. At step 2106, it is decided if the determined particletype is of interest. For example, for a particle monitor at a vineyard,a particle identified as a grass pollen is not of interest and theparticle monitor moves on to collect and analyze further airborneparticles. However, if the vineyard particle monitor identifies theparticle as a pathogenic fungal botrytis spore, the particle monitoringsystem moves on to step 2109. At step 2109, the particle monitoringsystem reaches a conclusion about the state of the biological particle.

In agricultural applications, it is of particular interest is todetermine if the biological particle, such as a pathological fungalspore, is in a healthy virulent state or in an injured and sterilestate. Other aspects of a biological particle's state may be ofinterest. For example, distinguishing between spores in a moist versusdesiccated states may aid predictions on time of (humidity activated)fungal growth from spores. Distinguishing between sexually and asexuallyproduced spores may provide information of interest relating to thatnature of a fungal reproduction and infestation. These are illustrationswhere not only the type, also but the state of biological airborneparticles is of interest to agriculture.

For non-agricultural applications such as personal pollen monitors,determinations of the state of airborne biological particles may also beof interest. The state of allergenic particles may well have an effecton a user's physiological reaction to exposure to a given number of theallergenic particles. For example, weakened UV fluorescence of pollengrains due to decomposition of fluorescing biomolecules with time orsunlight exposure may well correlate with decomposition of the pollengrain's allergenic biomolecules with time or sunlight exposure. Anotherexample of a non-agricultural application for the system and techniquesas discussed is the detection of airborne mold spores in support of moldinspection and mold remediation/removal from buildings. It is to beunderstood that the embodiments described below for agriculturalapplications illustrate principles that also apply to applications inother areas.

FIG. 21B shows a block diagram of a particle monitoring device 2120according to a specific embodiment. The particle monitoring device shownin FIG. 21B is similar to the particle monitoring device shown in FIG.3. The particle monitoring device shown in FIG. 21B, however, includes aspore state discrimination subsystem 2122. The state discriminationsubsystem includes a state discrimination manager 2124, state analysisengine 2126, reporting and log handler 2128, and a reference database2134 storing color characteristics of biological particles correspondingto illumination under various UV light. It should be appreciated thatthe blocks shown can be functional rather than structural so that it ispossible to have many different hardware configurations that can performthe illustrated functions. Implementation of the functional entities mayvary.

The state discrimination manager provides overall management of theprocesses to identify a state of a particle such as a fungal spore. Inparticular, the state discrimination manager is responsible fordirecting the operation of collection media motor 2136, opticalsubsystem 2138, and illumination subsystem 2140 to capture images ofparticles trapped within removable particle collection media 2142. In anembodiment, the collection media motor advances tape of the collectionmedia having the trapped particles to a position underneath the opticalsubsystem. The illumination subsystem illuminates a portion of the tapehaving the trapped particles with various light including visible lightand UV light. The optical subsystem captures images of the particleswhile the particles are being illuminated under each of the differentlighting conditions.

The UV light sources include light sources capable of generating UVlight of various spectral characteristics. For example, a first UV lightsource may provide UV light having a first spectral characteristic. Asecond UV light source may provide UV light having a second spectralcharacteristic, different from the first spectral characteristic. Athird UV light source may provide UV light having a third spectralcharacteristic, different from the first and second spectralcharacteristics. And so forth.

The images may be stored in an image repository 2144. The discriminationmanager retrieves the images from the image repository and passes theimages to the state analysis engine for analysis. The colorcharacteristics reference library stores a set of predetermined colorcharacteristics of fluorescent light for various types of fluorescentbiomolecules. In a specific embodiment, a predetermined colorcharacteristic of fluorescent light for a fluorescent biomolecule ofinterest corresponds to a concentration of the biomolecule of interestin a fungal spore of a known state.

In a specific embodiment, these color characteristics of fluorescentlight are associated with the fluorescent properties or attributes offluorescent biomolecules of interest. Each predetermined colorcharacteristic corresponds to excitement of a particular biomolecule ofinterest under UV light of a particular spectral characteristic.

For example, there can be a first color characteristic of fluorescentlight for a first biomolecule of interest corresponding to the firstbiomolecule of interest being illuminated under UV light having thefirst spectral characteristic. The first biomolecule may be in a fungalspore of a known state (e.g., virulent or sterile). Thus, the colorcharacteristic can be associated with the fungal spore of the knownstate.

Likewise, there can be a second color characteristic of fluorescentlight for a second biomolecule of interest corresponding to the secondbiomolecule of interest being illuminated under UV light having thefirst spectral characteristic. There can be a third color characteristicof fluorescent light for a third biomolecule of interest correspondingto the third biomolecule of interest being illuminated under UV lighthaving the first spectral characteristic. And so forth.

Likewise, there can be a first color characteristic of fluorescent lightfor the first biomolecule of interest corresponding to the firstbiomolecule of interest being illuminated under UV light having thesecond spectral characteristic, different from the first spectralcharacteristic. There can be a second color characteristic offluorescent light for the second biomolecule of interest correspondingto the second biomolecule of interest being illuminated under UV lighthaving the second spectral characteristic. There can be a third colorcharacteristic of fluorescent light for the third biomolecule ofinterest corresponding to the third biomolecule of interest beingilluminated under UV light having the second spectral characteristic.And so forth.

Likewise, there can be a first color characteristic of fluorescent lightfor the first biomolecule of interest corresponding to the firstbiomolecule of interest being illuminated under UV light having thethird spectral characteristic, different from the first and secondspectral characteristic. There can be a second color characteristic offluorescent light for the second biomolecule of interest correspondingto the second biomolecule of interest being illuminated under UV lighthaving the third spectral characteristic. There can be a third colorcharacteristic of fluorescent light for the third biomolecule ofinterest corresponding to the third biomolecule of interest beingilluminated under UV light having the third spectral characteristic. Andso forth.

Thus, the reference library stores predetermined color characteristicsof fluorescent light of various biomolecules of interest present in afungal spore of a known state (e.g., virulent or sterile). The colorcharacteristics correspond to the biomolecules of interest beingilluminated with UV light of various spectral characteristics. Eachpredetermined color characteristic may be tagged with a first tagidentifying a particular biomolecule of interest (e.g., flavin, NADH, ortryptophan), a second tag identifying a state of a fungal spore in whichthe particular biomolecule of interest is present, and a third tagidentifying UV light of a particular spectral characteristic that wasused to illuminate the fungal spore including the particular biomoleculeof interest.

In an embodiment, the optical subsystem generates a color image of atrapped particle while the trapped particle is illuminated by theillumination subsystem with UV light of a particular spectralcharacteristic. The state analysis engine determines a state of abiological particle (e.g., fungal spore) by measuring from the colorimage a degree and color of fluorescence for each pixel within anoutline of a fungal spore identified in the image. The analysis enginecompares the measurements against the color characteristics referencelibrary to identify which predetermined color characteristic offluorescent light most closely matches the measurements. Uponidentifying a matching predetermined color characteristic, across-reference can then be performed to the tag associated with thematching predetermined color characteristic storing the knowncorresponding state information.

Upon identifying the state, the reporting and log handler creates anentry in a log file. The entry may include a first field storing anidentification of the particle (e.g., powdery mildew), a second fieldstoring a timestamp indicating the time and date of particle capture,and a third field storing a determined state of the particle (e.g.,virulent or sterile).

The results of the state determination may further be displayed on auser interface 2146. The user interface may include an electronic screenon which the results are displayed. Instead or additionally, the resultsmay be transmitted to a client device (e.g., smartphone) 2148, remotecloud server 2150, or both. The transmission of the results may includea transmission of the images taken of the trapped particle. Instead oradditionally, the particle monitor may include a port (e.g., USB port)2152 in which the results, images, or both may be copied to a portableUSB drive plugged into the port. For example, in some cases, there maynot be network connectivity at the location where the particle monitordevice has been deployed. In this case, the user can download theresults, images, or both by traveling to the location of the particlemonitor and connecting the particle monitor to a portable drive (e.g.,flash drive).

FIGS. 21C-D show examples of results of the state determination that maybe output by the particle monitor device. In particular, FIG. 21C showsan electronic screen 2172 identifying a particle detected (e.g., powderymildew), a date and time of capture (e.g., Jul. 4, 2017), and adetermined state (e.g., virulent). FIG. 21D is similar to FIG. 21C. InFIG. 21D, however, another powdery mildew particle has been captured ata later date (e.g., Jul. 6, 2017) and its state has now been determinedto be sterile. As discussed, the state determination results may bedisplayed on an electronic screen of the particle monitor device,written to a log file at the particle monitor device, transmitted fromthe particle monitor device to a client device (e.g., smartphone),transmitted from the particle monitor device to a central server, orcombinations of these.

FIG. 22 shows a flow chart of a process for determining a state orhealth of a particle such as a fungal spore. FIGS. 23-26 schematicallyillustrate images of fungal spores of interest to agriculturalapplications. In some cases, a fungal spore of interest is an undesiredpathogen, but not necessarily. As a counter example, vitality ofairborne fungal spores of a highly sought after gourmet truffle may beconsidered to be a good thing. In any case, it is of interest to knowthe health state of spores in order to anticipate their ability toinitiate new fungal growths.

Referring now to FIG. 22, in a step 2210, a flow of air that may includea fungal spore is directed to a collection cartridge removably insertedinto a spore monitor. In a step 2215, the fungal spore is trapped withinor inside the collection cartridge. For example, the fungal spore may betrapped by an adhesive tape housed within the collection cartridge.

After the fungal spore has been trapped, the spore monitor advances aportion of the tape having the trapped fungal spore to the inspectionzone. In a step 2220, the fungal spore is illuminated with visible light(e.g., wavelengths ranging from about 390 nm to about 700 nm. In a step2225, while the fungal spore is illuminated with the visible light, thecamera sensor of the spore monitor captures a first image of the fungalspore.

FIG. 23 illustrates an image 2300 of a fungal spore illuminated byvisible light such as white light. Such a visible light image can beused by the spore monitor to determine the shape and texture of theexterior fungal spore wall 2320, thus providing morphologicalinformation for use in the identification of the spore type. The visiblelight image may provide a clear determination of a fungal sporesilhouette or outline 2310. In other words, in an embodiment, the sporemonitor analyzes the first image to identify or discern an outline ofthe fungal spore (step 2230—FIG. 22).

In a step 2235, the fungal spore is illuminated with ultraviolet (UV)light. In a step 2240, while the fungal spore is illuminated with UVlight, the camera sensor of the spore monitor captures a second image ofthe fungal spore. In a step 2245, the second image is analyzed tomeasure an intensity, degree, color, level of fluorescence, orcombinations of these within the outline of the fungal spore.

In a step 2250, based on the intensity of fluorescence (or othermeasured attributes or combination of measured attributes), a state ofthe fungal spore is identified. In other words, the intensity offluorescence can be used to determine the state or health of the trappedfungal spore. In an embodiment, there can be a predetermined thresholdintensity. The measured intensity is compared to the predeterminedthreshold intensity. If the measured intensity is above the threshold, adetermination may be made that the fungal spore is in a first state. Ifthe measured intensity is below the threshold, a determination may bemade that the fungal spore is in a second state, different from thefirst state.

For example, FIG. 24A illustrates an image 2400 of the UV fluorescencefor a healthy fungal spore. Within the fungal spore, the density offluorescing biomolecules may be different between the cytosol 2430 anddifferent types of cell organelles, with certain types of organelleshaving the highest densities of fluorescing biomolecules. (Cytosol isthe matrix fluid of the cell within the cell wall and within which arethe cell's organelles.)

To take a specific example, consider 365 nm UV illumination resulting influorescence mainly of flavins and also a contribution from NADH(nicotinamide adenine dinucleotide). Flavins and NADH play a key role incell respiration. The cell organelles known as mitochondria provideenergy to other parts of the cell by consuming fuel such as glucose andenergizing ATP molecules that then diffuse to other parts of the cell.(A fungal spore is a cell.) Flavins and NADH play a key role inenergizing of ATP molecules and hence flavins and NADH may be expectedto be present in higher concentrations at mitochondria. In anembodiment, a particle monitor includes a UV illumination sourceconfigured to generate and emit light at a wavelength of 365 nm. With365 nm UV illumination, mitochondria may be the strongly fluorescingcell organelles 2440 and the cytosol 2430 may fluoresce relativelyweakly within the fungal spore outline 2410.

For readers not familiar with the roles of mitochondria, flavins, NADHand ATP in cellular metabolism, the following analogy may beinformative. Imagine a city whose energy needs are provided by a fossilfuel power plant. The power plant of the city is analogous to themitochondria of a cell such as a fungal spore. Both the city's powerplant and the cell's mitochondria use hydrocarbons (e.g., fossil fuel orglucose) as an energy source. The heat-driven turbines of the city'spower plant are analogous to the “Krebs” or “citric-acid” cycle ofbiochemical reactions within the mitochondria. In analogy to energy fromthe city's power plant being delivered to homes of residents viaelectrical power lines, energy containing ATP molecules diffuse from themitochondria through the cytosol to various organelles and otherlocations within the cell.

In other words, ATP molecules go to various locations including thecells outer membrane and the cytosol itself. At the city's power plant,an electric generator plays an essential role transferring energy fromfossil-fuel powered turbines to the electrical power grid of the city.Flavins and the biomolecule NADH plays a role in the cell's energysystem that is analogous to the electric generator of the city's powerplant; flavins and NADH provide an essential energy-transfer linkbetween the biochemical reactions of the Krebs or citric-acid and theenergy transporting ATP biomolecules. Flavins and NADH are crucial tothe cellular energy system. Flavins and NADH are essential to sporemetabolism including respiration.

“NADH” is the higher energy form of the biomolecule “nicotinamideadenine dinucleotide” where the “H” refers to a hydrogen atom. If thehydrogen atom is removed, the result is the lower energy “NAD⁺” form ofnicotinamide adenine dinucleotide. The Krebs cycle “charges up”nicotinamide adenine dinucleotide molecules by converting NAD⁺ intoNADH. When NADH in turn “charges up” an energy carrying ATP molecule (byconverting ADP to ATP), it reverts to its lower energy NAD⁺ form.

These biochemistry details are of interest to biological particlemonitoring devices as described herein because only the higher-energyNADH form of nicotinamide adenine dinucleotide fluoresces whenilluminated by 365 nm ultraviolet light. The lower-energy NAD⁺ form ofnicotinamide adenine dinucleotide does not fluoresce. Hence theintensity of 365 nm UV fluorescence images of fungal spores is sensitiveto the energy charge state of the nicotinamide adenine dinucleotide. Ifthe respiration, including the Krebs cycle, of a fungal spore is shutdown by a fungicide, spore mitochondria will no longer be able toconvert lower-energy NAD⁺ to higher-energy NADH and hence UVfluorescence due to NADH will fade.

Interestingly, in contrast to NADH, the fluorescing form of flavins is alower-energy form of flavins while the “charged up” form of flavins doesnot fluoresce. Hence when mitochondria are actively metabolizing andproducing energy for the spore, we may expect a higher ratio of NADHfluorescence to flavin fluorescence relative to a spore in a dormantstate. Conversely, when mitochondria are not actively metabolizing andproducing energy for the spore, a higher ratio of flavin fluorescence toNADH fluorescence relative to the spore in an active or charged statemay be observed.

FIGS. 24B and 24C show images of mold material including spores. Theimage of FIG. 24B was captured shortly after the mold material wascaptured. The image of FIG. 24C is of the same mold material, butcaptured 15 hours later. Images of two mold spores are inside the solidcircles of FIG. 24B and the same spores are also circled in FIG. 24C.The spores seen in these figures are about thirty microns long. Much ofthe other mold material in the images are “hyphae.” Hyphae of mold areroughly analogous to roots or branches of plants. Hyphae are often inthe form of long thin lines, such as the hyphae indicated by the dashedcircle in FIGS. 24B and 24C. In this 15-hour interval, there was achange in the metabolic state of the imaged spores (and other mold partssuch as hyphae).

In this experiment, a 340 nm UV LED source of the device was on for theentire duration of the test, running at 100 mA. And the laboratoryambient temperature ranged from about 70 to 75 degrees per day, whilethe humidity ranged from about 35 percent to 40 percent.

The images show that mold spores will change color over time as they ageand their metabolic function changes. In particular, the color changesfrom a strong (high fluorescent) turquoise color to a more greenishcolor over time, such as over the 15-hour interval between the images ofFIGS. 24B and 24C. In some cases, the morphology of the spore may changewith time, such as becoming more long and narrow due to desiccation.

The particles captured by the cartridge adhesive of the prototypemonitoring device included brand new healthy spores feeding from a grapebunch. In this experiment, the cartridge was removed from theparticle-monitoring device, the adhesive touched on mold growing on thegraph bunch, and then the cartridge reinserted into theparticle-monitoring device. The images of FIGS. 24B and 24C are aportion of the field of view captured by the camera sensor. The grapebunch was located in a vineyard in Napa, Calif. known to have powderymildew.

In this specific embodiment, a spore state is determined by employing a340 nm UV excitation source in the monitoring device. A very healthy andyoung spore will have a turquoise color and a high degree offluorescence (intensity) when captured using an imager of the monitoringdevice. With time, the fluorescence color may change (e.g., to green)and the degree of fluorescence (intensity) may fade. The morphology orshapes of the fluorescent light images may also vary as the state of thespore changes. The use of a fungicide will accelerate the changes in thespore state within a few hours of being mortally wounded.

The environment plays a big role in the life of a spore. As an example,grape powdery mildew spores can live up to 60 days in a perfectenvironment that helps it thrive and then die of age. Certain fungicidesmay “kill” a spore within minutes while others may disable itsreproductive ability, thus altering their metabolic functions. An olderspore having very low metabolic function will die faster than a youngone when wounded by fungicides.

For a UV excitation wavelength of 340 nm (between the 325 nm and 365 nmexamples given by the dot-dot-dashed lines in FIG. 28S), NADH andriboflavin are the only fluorescing biomolecules that we are expecting.A 340 nm wavelength is too long (photon energy too low) to excitetryptophan in proteins or any other biomolecule that fluoresces inresponse to shorter UV wavelengths. NADH only fluoresces in itshigh-energy state (the lower-energy NAD+ state does not fluoresce) whileriboflavin is the opposite in that only its lower-energy statefluoresces. Neither molecule will fluoresce after they have decomposed.Furthermore, as seen in FIG. 28S, the higher-energy form of NADH emitsblue (about 455 nm) fluorescent light while the lower-energy form ofriboflavin emits green (about 520 nm) fluorescent light. Given thisscientific information, the following mapping between UV fluorescencebehavior and metabolic state may be expected to apply in many cases.Depending on the application different mappings may apply.

Blue (Turquoise) fluorescence may indicate a metabolically active state.Metabolic activity will put a significant fraction of NADH/NAD+molecules in their higher-energy “NADH” state in which they are holdinga hydrogen “H” ready for use in metabolic reactions; hence thismetabolically active state will emit blue fluorescent light. Much of theriboflavin will also be in a higher-energy state and hence notfluorescing but some riboflavin will remain in the lower-energy stateand fluoresce green light; this green light component in combinationwith the NADH blue may well explain the turquoise color seen in thedata. The blue (turquoise) fluorescent color is a signature of ametabolically active state. A metabolically active state may well be anindication of a healthy and freshly made spore.

Strong green fluorescence may indicate a metabolically resting state. Ahealthy spore that is resting or hibernating will contain a fullcomplement of NADH and riboflavin molecules. These molecules willgenerally be in their lower-energy state. In its lower-energy NAD+state, NADH does not fluoresce thus removing its blue light from thecamera sensor image. In contrast, the riboflavin molecules willfluoresce strongly precisely because they are in their lower-energystate. In this manner a strong green image is a signature of intactbiomolecules in a metabolically resting state. A metabolically restingstate may well be an indication of a healthy spore that is “hibernating”while it waits for conditions supportive of mold growth. However, if theright conditions never arrive, the metabolically resting state may alsoan indication of a spore under siege by unfavorable conditions and thatwill eventually lead to its death.

Weak green fluorescence may indicate a decomposing state. A spore thatis dying and decomposing will not only lack a blue NADH fluorescencesignature, but will have an increasingly dim green fluorescencesignatures as more and more (lower-energy) riboflavin moleculesdecompose.

When flavins fluoresce in response to UV light illumination, they emitlight at a wavelength of about 520 nm (green). When NADH fluoresces inresponse to UV light illumination, it emits light at a wavelength ofabout 460 nm (blue). Depending on the properties of the optical imagesensor, fluorescent light at 520 nm or 460 nm will produce distinctiveRGB color ratios. For example, for an image sensor with colorsensitivities as illustrated in FIG. 16, the ratio of intensitiesmeasured in red, green and blue sub-pixels will be approximateRED:GREEN:BLUE=1:10:4 for flavins and 0:1:8 for NADH. If measured RGBcolor ratios are approximately 1:10:4, the color ratios would support aninterpretation that the observed fluorescence is due to flavins withlittle contribution from NADH. However if another color ratio isobserved, then NADH and/or other biomolecules are contributing to theobserved fluorescence image. Such an ability to probe the biochemistrywith real-time optical image processing contributes to the ability of abiological particle monitor to probe the state as well as identity ofbiological particles under inspection.

FIG. 25 schematically illustrates an image 2500 of the UV fluorescenceof a fungal spore in which the intensity of fluorescence is weakcompared to that illustrated in FIG. 24A. Such an image would indicate alower density of fluorescing biomolecules in cytosol 2530 within fungalspore outline 2510 relative to cytosol 2430 as well as a lower densityof fluorescing biomolecules in cell organelles 2540 relative to cellorganelles 2440. If the illumination is 325 nm UV and the fluorescingbiomolecule is NADH, then an image such as that illustrated in FIG. 25would indicate a relatively low density of NADH and hence a relativelylow energy production within the cell, that is a low level ofrespiration.

Schematically illustrated image 2600 of FIG. 26 is similar to FIG. 24Aexcept for the presence of dark zones 2650 within the fungal sporeoutline 2610 where little or no fluorescence occurs. (For purposes ofillustration, FIGS. 23 through 27 are shown as negative images in thesense that the dark shading in the figures indicate an increase UVfluorescence and the white or absence of shading indicates no UVfluorescence.) In this case, under UV illumination, some of the cytosol2630, and perhaps some cell organelles 2640, goes dark. Such an imagewould suggest that certain local portions of the spore are ailing.

While not explicitly shown in a figure, it is also of interest toobserve when the effects illustrated in FIGS. 25 and 26 occur at thesame time. That is, when there are not only darkened local regions ofreduced UV fluorescence, but also an overall reduction in UVfluorescence intensity in the remaining regions of the spore.

FIG. 27 illustrates some of the modes of action of fungicides.Mitochondria disrupting fungicides 2760, such as Qol, cyazofamid,boscalid and flutolanil interfere with the function of mitochondria2740. A fungal spore damaged by such a mitochondria-disrupting fungicidemay well have low levels of flavins and NADH such as illustrated in FIG.25.

Cell wall disrupting fungicides 2770, such as polyoxin D, damage thecell wall 2710. Membrane disrupting fungicides 2780, such as DMI,dicarboximides, fludioximides, PCNB, chloroneb and propamocarb, damagecell membranes such as that immediately inside of the cell wall. Afungal spore damaged by either a wall disrupting fungicide or a membranedisrupting fungicides may suffer local damage where there is a breach inthe cell wall or membrane. This may result in UV fluorescence imagessuch as represented by FIG. 26. Analysis of visible light images (todetermine the location of the fungal spore outline) and at least oneUV-excited fluorescence image provides information on the state of afungal spore, such as whether it remains healthy and virulent or whetherit is injured, for example, by a fungicide.

As discussed above in connection with curve 1962 of FIG. 19, anultraviolet illumination source with a wavelength somewhat shorter than365 nm, such as 280 nm, will excite fluorescence of tryptophan and anyother fluorescent amino acids within proteins. In other words, the aminoacid tryptophan is excited by 280 nm UV light. There can be even shorterwavelengths that may excite other amino acids. Subtracting out the 365nm fluorescence image, the 280 nm excited fluorescence image willprovide image on the distribution of proteins within a fungal spore.Again this may well be used not only to aid biological particleidentification, but also determination of the state of the biologicalparticle. Similarly, even shorter UV wavelengths may be used tofluorescently excite even large sets of biomolecules, providing furtherinformation regarding both the identity and state of biologicalparticles of interest.

Once a particle-monitoring device has captured visible-light and UVfluorescence images, a number of options exist for extracting particlestate information from the image data. In an embodiment, explicitmathematical algorithms based on scientific considerations may beprovided based on quantitative measures of fluorescence intensity (e.g.,FIG. 25 versus FIG. 24A) and fluorescence non-uniformity (e.g., FIG. 26versus FIG. 24A). For example, a threshold of total intensity may bedefined below which a fungal spore is considered to be in an unhealthystate. Alternatively, machine-learning algorithms, such as often usedwith deep neural networks in the field of Artificial Intelligence (AI),may be used instead of, or in addition to, explicit scientifically basedmathematical algorithms.

In either case, there is great value in creating a learning set of data.A learning set of data may include, for example, visible-light and UVfluorescence images of a number of healthy and virulent fungal spores, anumber of fungal spores treated with a fungicide whose mode of action isto disrupt mitochondria (see item 2760 of FIG. 27), a number of fungalspores treated with a fungicide whose mode of action is to disrupt cellwalls (see item 2770 of FIG. 27), as well as a number of fungal sporestreated with a fungicide whose mode of action is to disrupt cellmembranes (see item 2780 of FIG. 27). Once provided with a sufficientlyinformative learning set of images, systems and techniques may beapplied to develop algorithms that distinguish between different statesof biological particles.

The ability to develop effective algorithms that distinguish betweendifferent states of biological particles is further enhanced by the useof a tape medium such as illustrated in FIGS. 8, 9 and 10. Having aphysical archive of captured biological particles such as fungal spores,particles of interest may be further studied off-line in a laboratory.For example, fungal spores may be removed from the tape medium andplaced in an appropriate nutrient and then incubated under appropriatetemperature and humidity conditions. If the spores grow into fungalcolonies, then one has the gold-standard of proof that the collectedfungal spores were virulent. If algorithms processing images predictedotherwise, then effort can be directed to fine tuning algorithmsaccordingly. If fungal colonies do not form, that would be consistentwith a prediction from imaging processing algorithms that inspectedfungal spores were not virulent.

However care must be given to the possibility that spore virulence waslost during storage in the tape medium. The tape medium may be stored incontrolled temperature and humidity conditions in order to preservespore vitality. In addition to attempted growth of fungal colonies,laboratory tests of fungal spores may include a number of bio-assaytests. In general, the option to supplement real-time optical imageprocess with laboratory study of archived physical samples of particlesprovides an import technique to test and improve the reliability of theresults of optical image processing.

Referring back now to FIG. 22, in a step 2255, the spore monitorcalculates a degree of confidence in the state assessment. If the degreeof confidence is above a threshold degree (e.g., state assessment issatisfactory), the spore monitor transmits and logs the state result(step 2260). Alternatively, if the degree of confidence is below thethreshold degree (e.g., state assessment is not satisfactory), the sporemonitor requests, obtains, and analyzes context information (step 2265).In a step 2270, the spore monitor again calculates a degree ofconfidence in the state assessment. If the degree of confidence is abovethe threshold (e.g., state assessment is satisfactory), the sporemonitor transmits and logs the state result (step 2260). If the degreeof confidence is below the threshold (e.g., state assessment is notsatisfactory), the spore monitor issues a request for review by a humantechnician (step 2275).

It should be appreciated that any number of images may be captured andthe images may be captured in any order. For example, the UV illuminatedimage may be captured before the visible light illuminated image. Thatis, the visible light illuminated image may be captured after the UVilluminated image. In some embodiments, only UV illuminated images arecaptured. That is, one can imagine situations in which no visible lightimage is captured. For example, if a protein fluorescence image turnsout to provide a better outline of the spore than the visible lightimage.

FIG. 28A shows a block diagram of a top view of a lighting orillumination arrangement of a particle monitor device according toanother specific embodiment. As shown in the example of FIG. 28A,particles 2804 trapped on a portion of tape 2806 have been positionedwithin a field of view 2808 of a camera sensor. The field of view isshown in the figure as a broken line. The particles may include fungalspores.

The illumination system of this monitoring device includes a first whitelight source or structure 2810A, a second white light source orstructure 2810B, a third white light source or structure 2810C, and anultraviolet light source or structure 2810D. In an embodiment, the lightsources include light emitting diodes (LEDs) or clusters of LEDs. Thelight sources are arranged about or around the field of view.

The light sources have illumination directions corresponding to thecorners of the field of view. More specifically, the first white lightsource is positioned so that its light arrives in the field of view froma direction corresponding to a first corner 2810E of the field of view.The distance from the white light LED 2810A to first corner 2810E islarger than shown in FIG. 28A. The sketch is not to scale in order toavoid drawing items too small to be clearly seen (like is generally donein for drawings of the solar system). The second white light source ispositioned so that its light arrives in the field of view from adirection corresponding to a second corner 2810F of the field of view.The third white light source is positioned so that its light arrives inthe field of view from a direction corresponding to a third corner 2810Gof the field of view. The UV light source is positioned so that itslight arrives in the field of view from a direction corresponding to afourth corner 2810H of the field of view. The dimensions of the field ofview, such as the distance from first corner 2810E and second corner2810F may be approximately one millimeter. The distance of the lightsources 2810A, 2810B, 2810C and 2810D from the field of view 2808 may beone or more centimeters.

The positioning of the light sources allow for illuminating the trappedparticles from different angles. In a specific embodiment, the lightsources are distributed and spaced equally about the field of view. Thelight sources may form corners of a square. A first distance is betweenfirst and second white light sources. A second distance is betweensecond and third white light sources. A third distance is between thethird white light source and the UV light source. A fourth distance isbetween the UV light source and the first white light source. In thisspecific embodiment, the distances are equal to each other. In anotherspecific embodiment, at least one distance is different from anotherdistance. For example, the light sources may form corners of a rectangleor other polygon. It should be appreciated that the light sources shownin FIG. 28A and elsewhere can represent openings through which light isoutput towards the field of view. For example, an opening may include anend of an optical fiber. An opposite end of the optical fiber may beconnected to the actual emitter (e.g., LED) that may be locatedelsewhere in the particle monitor device and remote from the field ofview. The optical fiber guides or transmits light from the actualemitter to the opening near the field of view.

In an embodiment, a technique for identifying a spore uses bursts orflashes of white light in combination with illuminating the trappedparticles with UV light. Before the health of a fungal spore can bedetermined, an identification of the type of spore it is may be made.One of the key parameters employed in identifying these is themorphology of the particle among others.

Fungal spores are in many cases transparent or semi-transparent undervisible light, which presents a challenge in obtaining morphology usingwhite light, as most of it will go through it. These are alsopleomorphic where their size and shape can vary over time, that is,spores are like frogs that also change shape during their lifecycle(think tadpole versus adult frog). Desiccation or moisture absorptionmay also cause spore shape changes. Typical techniques used today employlight microscope, SEM, AFM microscopy, among others; and are timeconsuming, requiring sample preparation and trained users of the deviceand the technique employed.

It can be cumbersome and time consuming when viewing a transparentfungal spore such as Erysiphe Necator (aka. Powdery mildew) or Botrytis(aka. Gray mold) for a user to apply a staining dye to enhance theoutline of the spore in order to determine the shape (morphology) of thespore using a camera or estimating it against a size grid. In anotherexample, when using a fluorescent microscope it is necessary to applyfluorescent dyes to the subject and further manipulate the emitted lightthrough beam splitters and filters. Typically light is polarized and, orcollimated. AFM techniques may employ force, beam deflection, contact,phase measurements, electrical fields, magnetic fields, and others tomap out the shape of a particle.

In order to obtain the shape of a spore or simultaneously scan a groupof spores without doing any sample preparation, applicant has discoveredthat while exciting the spore(s) with a UVLED (non-polarized) andapplying a burst of white light from a non-polarized light source at adifferent angle from the UVLED excitation angle, near the end of theintegration time results in a single image that contains fluorescentdata as well as a profile of the full area (shape) of the particle.

Applicant has discovered that due to the pleomorphic nature of thefungal spores and that they do not always come in front of the cameralens in the same direction and their health state, it is desirable toinput white light in smaller bursts around the perimeter of theparticle.

FIG. 28B shows an overall flow of the white light burst or flashtechnique according to a specific embodiment. In a step 2820A, a portionof tape having a trapped particle is positioned within a field of viewof a camera sensor of the particle monitor. In a step 2820B, a UV lightsource (e.g., UV LED) is activated to illuminate the trapped particlewith UV light. The activation of the UV light source is accompanied bythe opening of a camera shutter associated with the camera sensor (step2820C). Optionally, shutter opening of step 2820C may occursimultaneously with, or even before, the UV light activation of step2820B. In a step 2820D, one or more bursts of white light is directedtowards the trapped particles within the field of view. In a step 2820E,the camera sensor is closed to generate an image. In a step 2820F, theimage is analyzed to obtain a shape of the trapped particle.

FIGS. 28C-28E show a comparison of various image techniques on fungalpowdery mildew. FIG. 28C shows an image of the fungal as illuminatedunder white light. FIG. 28D shows an image of the fungal as illuminatedunder UV light. FIG. 28E shows an image of the fungal as illuminatedunder UV light and a burst of white light.

More particularly, in this experiment, powdery mildew spores along withconidia and some hyphae were imaged using white light (FIG. 28C), a 340nm UV light (FIG. 28D), and the UV plus white light burst (FIG. 28E). Aduration of the white light burst was 0.1 seconds. Based on the imagingresults, UV plus the white light burst was shown to provide betterdefinition of total particle area and shape (used to extract importantclassification parameters) along with UV fluorescence used to determinethe health state of the fungal spore. In this experiment, the whitelight image was collected much earlier than the UV and UV plus whiteburst images and the particles in the field of view had shifted to theleft by approximately 0.25 mm.

FIG. 28F shows further detail of a flow for the white light burst orflash technique according to a specific embodiment. In a step 2835A, aportion of a tape having a trapped particle is positioned within a fieldof view of a camera sensor. In a step 2835B, a UV light source (e.g., UVLED) is activated to illuminate the trapped particle with UV light. In astep 2835C, a camera shutter associated with the camera sensor is openedfor a time period. The time period may be referred to as an exposuretime period.

In a step 2835D, while the trapped particle is illuminated with the UVlight, the camera sensor is allowed to collect light emitted from thetrapped particle during a first portion of the time period.

In a step 2835E, after the first portion of the time period has elapsed,past, or expired, a first burst of white light, originating from a firstposition, is directed during a second portion of the time period afterthe first portion towards the trapped particle.

In a step 2835F, after the first burst, a second burst of white light isdirected during the second portion of the time period towards thetrapped particle. The second burst of white light originates from asecond position, different from the first position.

In a step 2835G, after the first and second bursts, a third burst ofwhite light is directed during the second portion of the time periodtowards the trapped particle. The third burst of white light originatesfrom a third position, different from the first and second positions.

In a step 2835H, after the second portion of the time period haselapsed, the camera shutter is closed to generate an image.

In a step 2835I, the image is analyzed to obtain a shape of theparticle.

In a specific embodiment, a duration of the time period is about 15seconds, a duration of the first portion of the time period is about 14seconds, and a duration of the second portion of the time period isabout 1 second.

FIG. 28G shows a timeline and sequence of events of the white lightburst technique according to a specific embodiment. As shown in theexample of FIG. 28G, there is a time period or exposure time period2840A, first portion 2840B of the time period, and second portion 2840Cof the time period. The second portion is immediately after the firstportion. In other words, the first portion is immediately before thesecond portion. In this specific embodiment, durations of the first andsecond portions are different. In this specific embodiment, the durationof the first portion is greater than the duration of the second portion.In other words, the duration of the second portion is less than theduration of the first portion.

At T0, initial events 2840D of the first portion of the time periodincludes activating a UV LED and opening the camera shutter. The firstportion of the time period ends at T1 at which point the second portionof the time period begins. The ending of the first portion of the timeperiod (or the beginning of the second portion of the time period) isaccompanied by a first burst of white light 2840E. During the secondportion of the time period, there is a second burst of white light 2840Fat T2 that follows the first burst of white light. In other words, thesecond burst is after the first burst. The first burst is before thesecond burst.

During the second portion of the time period, there is a third burst ofwhite light 2840G at T3 that follows the second burst of white light. Inother words, the third burst is after the first and second bursts. Thefirst and second bursts are before the third burst. The second portionof the time period ends at T4 and is accompanied by a closing of thecamera shutter to generate an image 2840H.

In a specific embodiment, the UV light and white light bursts originatefrom different positions about the field of view of the camera sensor.For example, as shown in the block diagram of FIG. 28A, the lightsources or light openings are arranged to illuminate from differentdirections about or around the field of view. This allows various lightto be directed towards the trapped particles at various differentangles. In particular, the first white light source is closer to thefirst corner of the field of view than the second, third, and fourthcorners. The second white light source is closer to the second corner ofthe field of view than the first, third, or fourth corners. The thirdwhite light source is closer to the third corner of the field of viewthan the first, second, and fourth corners. The UV light source iscloser to the fourth corner of the field of view than the first, second,or third corners.

In a specific embodiment, a duration of the time period is 15 seconds, aduration of the first portion of the time period is 14 seconds, aduration of the second portion of the time period is 1 second, aduration of the first burst of white light is 0.033 seconds, a durationof the second burst of white light is 0.033 seconds, and a duration ofthe third burst of white light is 0.033 seconds. In a specificembodiment, a method includes first turning on the UVLED from the bottomright of the field of view and opening the camera shutter to collectemitted light for 15 seconds. At 14 seconds of completion, turning on awhite LED for 0.033 secs from the bottom left of the field of view, thenturning it off while turning on a second white LED coming from the topleft of the field of view for 0.033 secs, then turning it off whileturning on a third white LED in the top right of the field of view for atotal of 0.033 secs. The shutter then closes at 15 secs and the cameragenerates a single image showing fluorescence of the particles withwhite light enhancing the fluorescent light within the particle andfilling in the entire volume of a spore defined by its outline,perimeter, or boundary.

As shown in the timeline example of FIG. 28G, the white light burst isorders of magnitude smaller than the UV integration time. Applicant hasdiscovered that a tenth of a second (0.1 secs) is plenty of white lightover 15 secs of UV fluorescence, for at least for powdery mildew spores.Adding more white light integration time results in more imagesaturation, which causes finer spore details to be lost. And having toolittle does not add enough detail to the spore shape. Other spores maybenefit from a different setup.

In a specific embodiment, the white light burst occurs right before theend of the integration time. In this specific embodiment, applicant hasdiscovered that capturing these at beginning or middle of integrationtime does not provide enough detail when UV fluorescence continues to becaptured after white light was acquired.

This could be done at more angles and directions than just three and itcan be desirable to have these also alternate from different incidentangles above the tape surface. In a specific embodiment, there are atleast two white light sources and one UV light source at 120-degreesapart from each other.

FIGS. 28H-O show a series of intermediate sample images made of grapepowdery mildew under various lighting conditions or modes using thelighting arrangement shown in FIG. 28A. These intermediate sample imageshelp to illustrate the advantages and benefits of combining UV and whitelight bursts or flashes when imaging a trapped particle foridentification and analysis.

More specifically, FIG. 28H shows an image generated while UV light iscoming from bottom right of the field of view. FIG. 28I shows an imagegenerated while a direction of illumination was from bottom left of thefield of view. FIG. 28J shows an image generated while a direction ofillumination was from top left of the field of view. FIG. 28K shows animage generated while a direction of illumination was from top right ofthe field of view. FIG. 28L shows an image generated where all threewhite LEDs were on simultaneously for 1 second. FIG. 28M shows an imagegenerated where illumination included a single (white) LED and no UVlight was present. FIG. 28N shows an image where the white lightbursting or flashing occurred early during the exposure time period.FIG. 28O shows an image where the white light bursting or flashingoccurred late during the exposure time period.

FIG. 28P shows a top view of a lighting or illumination arrangement of aparticle monitor according to another specific embodiment. FIG. 28P issimilar to FIG. 28A. In FIG. 28P, however, the illumination systemincludes two white light sources (a first white light source 2860A, asecond white light source 2860B), and a UV light source 2860C. The lightsource direct, from different directions, light towards a field of view2860D of a camera sensor. As shown in the example of FIG. 28P, the lightsources are disposed about or around the field of view and are equallyspaced from each other. In other words, in this specific embodiment, thelight sources are positioned such that a circle 2860E passes through acenter of each light source. A first line segment 2860F extends from acenter of the circle to a center of the first white light source. Asecond line segment 2860G extends from the center of the circle to acenter of the second white light source. A third line segment 2860Hextends from the center of the circle to a center of the UV lightsource.

A first angle 2860I is between the first and second line segments. Asecond angle 2860J is between the second and third line segments. Athird angle 2860K is between the third and first line segments. In aspecific embodiment, the first, second, and third angles are the same.In a specific embodiment, each of the first, second, and third anglesare 120 degrees.

In an embodiment, non-polarized light is used so that light scatters inrandom angles and reflects randomly. This can help against the everchanging topography of the spores throughout their life cycle. Whenyoung their outer shell may have a soft surface and be full of moisturewhich is more reflecting, but under stress like loss of moisture it willwrinkle and shrink in size having varying areas of the spore that willreflect more or less than others. In contrast, having light coming fromall directions at once may fill in details that are not clearly visiblefrom certain illumination angles.

Some light goes through the particle, some may get tunneled (orchanneled) within the outer walls (the skin) of the spores, and somelight will bounce in different directions inside the spore as it goesthrough mitochondria, lipids, and others where absorbed versus reflectedlight will vary. In some cases, illuminating from a single angle may notreveal the entire shape, but illuminating from multiple angles atdifferent intervals reveals more finer details of the spores.

As the spores age, certain structures will be weaker than others andwill take a lot more energy to emit light.

In some cases, a single or multi-angle white light image may not provideenough spore shape details. The amount of reflected white light acrossthe spore may correspond to its health (age as well) as seen through theUV fluorescence. Newly released spores have very bright turquoisefluorescence and a very bright reflected white light across the entireshape of the spore. However, after days later the turquoise fluorescencedecreases or becomes green and applicant has discovered that whencollecting a single white light image of the same previous spore nowshows dimmer, less reflecting, or dark areas within the spore. This mayindicate that one cannot necessarily rely on a white light image toextract shape because it will also change with the spore's health.

Using UV fluorescence with the short burst of white light integratedfrom different angles and across different directions across the spores'camera field of view can provide an accurate rendition of the spores'volume. And instead, there can be a single fluorescent and single whitelight against the combined image to detect the spore state.

Below are some benefits of this collection scheme:

1) A single image contains accurate spore shape, which is one of manyparameters used in identification. Contains fluorescent informationacross its shape to determine spore state

2) Additional information about spore texture can be extracted tofurther identify a particle within its state.

3) A new set of color parameters can be extracted and used in sporeidentification at various stages of the life of the species.

In various specific embodiments, a new method and technique is providedfor extracting shape information of transparent and semi-transparentparticles including fungal spores and other organisms regardless of thestage in their lifecycle. In a specific embodiment, a method is providedof simultaneously generating shape, texture, and topography data of atransparent spore. In another specific embodiment, there is a system forsimultaneous discrimination of random particles and theirclassification.

FIG. 28P shows a symmetrical arrangement of the light sources. Invarious other specific embodiments, there may be a non-symmetrical orasymmetrical arrangement of the light sources. For example, one or moreof the first, second, or third angles may be different from another ofthe first, second, or third angles. In another specific embodiment, alight source may be in a plane different from a plane of another lightsource.

FIG. 28P shows an example of light sources whose positions are fixed.FIGS. 28Q and 28R show top views of another lighting arrangement of aparticle monitor device according to another specific embodiment. FIGS.28Q and 28R are similar to FIG. 28P. In FIGS. 28Q and 28R, however,there is a single white light source whose position is moveable. Moreparticularly, FIG. 28Q shows a lighting or illumination system thatincludes a white light source 2875A and a UV light source 2875B. In thisspecific embodiment, at least one of the light sources is moveablerelative to particles 2875C trapped on a cartridge collection tape2875D.

Specifically, FIG. 28Q shows the white light source in a first position2875E. FIG. 28R shows the white source having moved 2875F from the firstposition to a second position 2875G, different from the first position.The light source (e.g., white light source) can be moved so that theparticles can be illuminated from different angles and directions. In aspecific embodiment, one or more light sources are rotatable about thefield of view. A light source may rotate, translate, or both so as toilluminate particles within the field of view from different angles anddirections.

In a specific embodiment, illumination of the particle with LEDs is fromfixed angles about the tape where the spore is positioned under thecamera. In another specific embodiment, a device may include only asingle white light source and the light source can move about the sporeand illuminate it from different directions and angles.

In a specific embodiment, at least three positions or angles—120 degreesapart are provided to aid in generating a contour profile of the spore.There can be a ‘turret’ holding a single white and UV LEDs that rotatesaround the camera's field of view illuminating and the camera collectingimages from different illumination angles.

In another specific embodiment, there can be a stage holding the spore(the tape) can move from a known XY location to a known new XY locationand illuminated again where both image locations are overlap usingsoftware to now reveal an enhanced contour profile of the particle, thiscould be done multiple times until a satisfactory profile has beenobtained, which can be determined when a new XY location does not changethe previous area profile of the particle (spore). In another specificembodiment, the lights may be fixed but the camera moves about to allowfor images from different light angles.

As another example, a particle monitor device may include a multiplexer,computer controller and control circuit, light openings disposed indifferent positions, locations, or angles about a field of view of acamera sensor of the monitor, a white light emitter, and a UV lightemitter. At least a subset of the light openings may be connected to anoutput of the multiplexer. Another subset of the light openings may beconnected to the UV light emitter.

The multiplexer may be connected to the white light emitter. Forexample, first and second light openings may be connected via opticalfiber to the output of the multiplexer. The white light emitter may beconnected via optical fiber to the multiplexer. The first light openingmay be positioned about the field of view to output light in a firstdirection or angle towards the field of view. The second light openingmay be positioned about the field of view to output light in a seconddirection or angle, different from the first direction or angle, towardsthe field of view.

The computer controller and control circuit is interconnected with andcontrols the operation of the multiplexer, camera sensor, and emitters.The computer can use the multiplexer to control which of the first orsecond light openings to use to transmit light from the white lightemitter to the field of view.

For example, a first burst of white light may be transmitted from thewhite light emitter through the multiplexer and out the first lightopening towards the field of view. The first burst of white light willnot be transmitted through the second light opening because themultiplexer can prevent the transmission. A second burst of white lightmay be transmitted from the same white light emitter through themultiplexer and out the second light opening towards the field of view.The second burst of white light will not be transmitted through thefirst light opening because the multiplexer can prevent thetransmission.

Thus, a single light emitter can be used to output light in two or moredifferent directions or angles at different times. This can help toreduce the cost of the particle monitor. As one of skill in the art willrecognize, a UV light emitter can similarly be connected to amultiplexer to provide different UV illumination lighting at differenttimes, directions, or both. Alternatively, the multiplexer may beconfigured to provide illumination (e.g., white light, UV light, orboth) from different angles or directions simultaneously orconcurrently.

Current prior art devices include some sort of impactor or undesiredparticle purge prior to analyzing a set of particles of interest. In aspecific embodiment, a particle monitor device is provided that is notlimited to any particle size or shape, rather the flow of air isunobstructed, unrestricted, and unlimited in the particles that getanalyzed, and analysis is only limited by camera resolution. In aspecific embodiment, light illumination comes from at least twodirections to generate the contour profile of the particle (spore). In aspecific embodiment, white light is added as a burst to UV light ingenerating the contour profile of the particle (spore). In a specificembodiment, the spores are illuminated against a dark (black)background.

Conventional fluorescence spectroscopy takes advantage of the highspectral resolution of spectrometers such as those based on diffractiongratings. Such spectrometers can be very expensive. In comparison, thecolor resolution of a color camera sensor is very crude as is seen bythe broad curves of red, green and blue sub-pixels in the plot of FIG.16. The embodiments discussed in this application make use of colorcamera sensors due to their ability to provide rich morphology data atlow cost, even at the cost of loosing color spectral resolution.Nevertheless, surprisingly, the humble color resolution of camerasensors are still sufficient to extract useful color information withwhich to separate contributions of different types of fluorescentbiomolecules and hence probe the state of fungal spores. To furtherillustrate this point, a further detailed embodiment is discussed below.

Referring to FIG. 28S, consider fungal spores for which the dominantbiomolecules contributing to fluorescence images are riboflavin(understood to also include related biomolecules), NADH (also understoodto include related molecules including NADPH), and tryptophan (alsounderstood to include any other fluorescing amino acids). Suchfluorescing biomolecules may be excited by blue or ultraviolet (UV)light of wavelength λ_(EX) resulting in the emission of light ofwavelength λ_(EM). FIG. 28S represents the wavelength of colorcharacteristics of these three types of biomolecules where inside theheavy circles are values wavelength pairs (λ_(EX), λ_(EM)) of relativelystronger fluorescence and inside the finer lines relatively weaker butnon-zero fluorescence. Riboflavin has three particularly favorableexcitation wavelengths, all of which result in the same emissionwavelength.

In this detailed embodiment, four fluorescence images of fungal sporesare captured at excitation wavelengths of about 445 nm, 365 nm, 325 nmand 280 nm. These excitation wavelengths correspond to the dot-dot-dashhorizontal lines drawing in FIG. 28S. Corresponding biomoleculefluorescence wavelength conversions are illustrated in FIG. 29.Riboflavin results in emission at about 520 nm as indicated by items(a), (b), (d) and (f) of FIG. 29, while NADH results in emission atabout 455 nm in items (c) and (e). Only the shortest wavelengthexcitation 280 nm is able to excite tryptophan, and only thelonger-wavelength tail of the resulting emission spectrum is in thevisible spectrum and detectable by the camera sensor (according to FIG.16). In this example, only riboflavin contributes to fluorescence due to445 nm excitation. The fluorescence signal from 365 nm illumination isdue to both riboflavin and NADH with riboflavin dominating. Thefluorescence signal from 325 nm also is due to both riboflavin and NADH,but this time with NADH dominating. Finally the fluorescence signal from280 nm includes a contribution of tryptophan as well as riboflavin.

The distributions of flavins, NADH, and proteins (amino acid chains,most of contain at some level the amino acid tryptophan) provideimportant clues about the state of a spore. Let “C_(RIBOFLAVIN)(x,y)”represent the distribution or concentration of riboflavin and relatedbiomolecules as a function of the coordinates (x,y) of the camera sensorpixels within the outline of a fungal spore. Likewise let us define“C_(NADH)(x,y)” and “C_(TRYPTOPHAN)(x,y)” similarly for thedistributions of NADH and tryptophan. For example, the distributions“C_(RIBOFLAVIN)(x,y)” and “C_(NADH)(x,y)” may have peaks of highconcentration at the locations of mitochondria while the distribution“C_(TRYPTOPHAN)(x,y)” may have peaks where there are high concentrationsof proteins. The biomolecule distributions “C_(RIBOFLAVIN)(x,y)”,“C_(NADH)(x,y)” and “C_(TRYPTOPHAN)(x,y)” are of interest for assessingthe state of a fungal spore.

However, these biomolecule distributions of interest cannot be directlymeasured, but must be inferred from red, green and blue color pixel datafrom camera images. Let “RGB_(i)(x,y)” represent the signal from anRGB-camera-sensor color subpixel “i” (where i=RED, GREEN or BLUE) forthe pixel at location (x,y); a superscript may be added to explicitlydefine the excitation wavelength λ_(EX). FIG. 30 provides a generalformula for relating measured pixel values “RGB_(i)(x,y)” to thebiomolecule distributions “C_(j)(x,y)” where j=RIBOFLAVIN, NADH orTRYPTOPHAN. The quantum efficiencies of the color sub-pixels as afunction of wavelength, such as plotted in FIG. 16, are represented by“p_(i)(λ)” where again i=RED, GREEN or BLUE. The fluorescencecharacteristics of the biomolecules, such as plotted in FIG. 28S, isrepresented by the factor “F_(j)” where a superscript “λ_(EX)→λ_(EM)”indicates a two-dimensional location (λ_(EX), λ_(EM)) in the plot ofFIG. 28S. In FIG. 31, the general formula of FIG. 30 is explicitlyapplied to the biomolecule fluorescences of FIG. 29.

From captured fluorescence images, the numerical values of left handsides of the equations of FIG. 31 are known. In the method of thisembodiment, the color sensitivity characteristics of the camera sensor,“p_(i)(λ)”, are predetermined via calibration procedures or othertechniques.

Similarly, laboratory studies of the fluorescence properties ofbiomolecules of interest, perhaps as reported in the publishedscientific literature, predetermine the values of “F_(j)” for sets ofwavelengths “λ_(EX)→λ_(EM)” of interest. As a result, for each pixelwithin the outline of a fungal spore, in FIG. 31, there are ten linearequations with three unknowns, namely the biomolecule distributions“C_(RIBOFLAVIN)(x,y)”, “C_(NADH)(x,y)” and “C_(TRYPTOPHAN)(x,y).” Hencethere is redundant information from which to compute the threebiomolecule distributions with the aid of well-known methods of solvingsets of linear equations. Thus, based on the equation of FIG. 30, it ispossible to determine biomolecule distributions from capturedfluorescence images.

In other words, as long as the number of independent equations isgreater than or equal to the number of biomolecule distributions to beextracted, it is feasible to do so. For example, even without the imagesfor 445 nm blue illumination and 325 nm UV illumination, thedistributions of the three types of biomolecules may still bedetermined. Similarly, if all four illumination wavelengths areretained, but several additional biomolecules are found to contribute tothe fluorescence images, the distributions of the additional types ofbiomolecules may also be determined.

Illumination wavelengths 365 nm, 325 nm and 280 nm are outside thevisible spectrum and well into the ultraviolet spectrum to which thecamera sensor does not respond. However, the longest illuminationwavelength 445 nm is still within the visible spectrum and correspondsto a blue color. Referring to FIG. 16, signals of blue camera sensorsub-pixels, and even green camera-sensor sub-pixels, will be dominatedby direct scattering of the blue 445 nm light making it difficult todetect the presence of relatively weaker 520 nm fluorescent light. The445 nm→520 nm fluorescence signal is most easily detected with the redcamera-sensor sub-pixels as red sub-pixels have some sensitivity togreen 520 nm fluorescent light (p_(RED)(520 nm)≠0) and at the same timethe red sub-pixels are very insensitive to the blue 445 nm illuminationlight (p_(RED)(445 nm)≈0). Red camera-sensor sub-pixels signals for 445nm illumination have the advantage of providing images of the flavindistribution unmixed with fluorescent signals of biomolecules excitedonly by ultraviolet light.

The above discussion corresponds to steps 3210, 3220, 3230 and 3240 ofthe flow chart of FIG. 32. In a specific embodiment, a method mayinclude capturing, by a camera sensor, an image of a fungal spore whilethe fungal spore is illuminated with UV light; inferring concentrationsof biomolecules of interest within the fungal spore by correlating,associating, or relating a value of a pixel located at coordinates (x,y)on the image to a concentration of a biomolecule of interest as being atthe coordinates (x,y) on the image; obtaining reference informationcomprising fluorescent properties of the biomolecules of interestassociated with known states of the fungal spore; obtaining camerasensor information comprising color sensitivity characteristics of thecamera sensor; and processing the inferred concentrations of thebiomolecules of interest within the fungal spore with the reference andcamera sensor information to identify a state of the fungal spore.

In applications in which skilled specialists are available to interpretthe data, it may be advantageous, as in step 3250, for the system tovisually present to a human operator the results of the computations ofbiomolecule distributions. In an embodiment, the process ends at step3250 and steps 3260 and 3270 are omitted. In this specific embodiment,the monitor does not proceed to determine the state of fungal spore, butrather leaves that to a human operator. Alternatively, the system mayhave appropriate software to provide automatic determination of thestate of the fungal spore based on the biomolecule distributions withinthe outline of the fungal spore.

Of particular interest in agricultural application is the differencebetween a viable spore of a fungal pathogen that is capable of spreadingdisease and a fatally injured or dead spore that is not capable ofspreading disease. That is, it is of interest to test the infectiousnessof detected pathogenic fungal spores. A conventional laboratory test ofspore virility is to incubate spores in a nutrient medium underappropriate temperature and humidity conditions and observe whether ornot the spores grow into visible mold colonies. While a convincing testof infectiousness, the time delay and labor costs of such a test isoften disadvantageous. Building on the method of FIG. 32, a much fasterand lower-cost test of fungal spore infectiousness is provided.

Referring to FIG. 33, step 3310 corresponds to steps 3210, 3220 and 3230of the flow chart of FIG. 32 and results in the capture of both visiblelight and fluorescent light images of a fungal spore. Step 3320 of FIG.33 corresponds to steps 3240 through 3270 in which the state of thefungal spore is determined. In particular, step 3320 determines a firststate of a capture fungal spore, namely the state of the spore shortlyafter being captured from ambient air.

In step 3330, the captured fungal spore is subjected to conditions thatencourage spore germination and growth. Conditions to encouragegermination may include, but are not limited to, humidity, temperature,and availability of nutrients. For example, via motion of the adhesivecoated tape within the cartridge, a spore sample may be moved to alocation of elevated humidity and temperature, and perhaps a small dropof sugar water wets the fungal spore. In other words, in a specificembodiment, a particle monitor includes an environmental regulator,device, or subsystem. The environmental regulator is responsible forproviding a controlled environment for the captured fungal spore inorder to assess the state of a captured fungal spore. The environmentalregulator may include, for example, a humidifier, dehumidifier, or bothto control or change a humidity level within the particle monitor; anair conditioner, heater, or both to control or change a temperaturewithin the particle monitor; a feeder to provide the fungal spore withnutrients, or combinations of these.

In step 3340, enough time is allowed to pass during which the fungalspore responds to germination inducing conditions with a change inmetabolic state. For example, a delay may be implemented between thesetting of the environmental conditions and capturing of a second image.It is important to note that the wait time of step 3340 is very briefcompared to the time it takes a spore to grown into a mold colony as inconventional viability tests. It may not even be necessary to wait forthe spore to split into multiple cells. To determine whether or not afungal spore has been killed or fatally injured by fungicide, it may besufficient to observe metabolic changes, or lack thereof, with a singlespore cell. For example, it may be sufficient to observe the degree ofincreased metabolic activity in the spore's mitochondria as determinedfrom the riboflavin and NADH fluorescence signals.

In step 3350, steps 3210, 3220 and 3230 of FIG. 32 are repeated in orderto capture second images of the fungal spore after it has been subjectedto germination inducing conditions. In step 3360, steps 3240 through3270 are repeated but this time with the second rather than firstcaptured set of images resulting in the determination of a secondpost-germination-condition state of the fungal spore. In step 3370, thesecond fungal spore state as determined in step 3360 is compared to thefirst fungal spore state as determined in step 3320 and a determinationis made of the infectiousness of the fungal spore captured from ambientair.

In an embodiment, a feature of the system analyzes both morphology andfluorescence properties of particles that have been trapped and imagedusing a color camera sensor. The combined analysis can be used to notonly identify a particle, but to also assess a state of the particle(e.g., healthy versus not healthy, active versus dormant, or aliveversus dead). The morphology analysis facilitates an identification ofthe various parts or anatomy of a cell (e.g., mitochondria versus cellwall). Consider, as an example, a fungicide designed to interrupt aspecific process of the cell or to target a specific portion of the cell(e.g., mitochondria). Since different parts of the cell may havediffering properties with respect to fluorescence, analyzing imagescaptured under different illumination conditions can be used todetermine whether or not the fungicide has been effective.

Determining a state of a fungal spore may include analyzing contextinformation. For example, if the context information indicates thatenvironmental conditions are conducive to germination and an imageanalysis indicates that the fungal spore is inactive, a determinationmay be made that the fungal spore is actually dead rather than in adormant or hibernating state. Conversely, if the context informationindicates that environmental conditions are not conducive to germinationand an image analysis indicates that the fungal spore is inactive, adetermination may be made that the fungal spore may be in a dormantstate and a further analysis is required to determine whether or not thespore is dead.

FIG. 34 illustrates the contents of an exemplary particle informationpacket 3400 that may be generated by particle monitor 105 in anembodiment, in connection with analyzing particles captured by theparticle monitor. FIG. 35 shows a particle information packet history.

Referring now to FIG. 34, shown are parameters of a data structurestoring characteristics of a particle and metadata associated with theparticle. The data structure may be stored in the memory or storage of aphysical computing device. The data structure may be implemented, forexample, as a table of a database. While FIG. 34 shows some specificexamples of particle characteristics and metadata that may be collected,derived, and stored for a particle, it should be appreciated that therecan be instead or additionally other particle characteristics,associated metadata, or both that may be stored.

Consider, as an example, a particle monitor placed in a vineyard tomonitor pathogenic fungal spores such as powdery mildew. In thiscontext, FIG. 35 illustrates an example hypothetical history of oneparticle information packet.

When a particle is observed in the field of view of the camera sensor ofthe particle monitor, a particle information packet 3400 (FIG. 34) iscreated (step 3505—FIG. 35). At creation, it includes a packet header3410 (FIG. 34) including particle ID number 3412 and pointers 3414 tovarious blocks in the packet, a particle ID block 3420 containing items3421 through 3426, as well as an objectives block 3430 with anapplication type 3432 of “agricultural monitoring” and a definition ofparticles of interest 3434 of “powdery mildew.”

In the example shown in FIG. 34, the particle ID block includes atimestamp 3421, particle collection device serial number 3422, deviceGPS coordinates 3423, adhesive-coated tape reel number 3424,x-coordinate of particle along length of tape 3425, and y-coordinate ofthe particle (perpendicular to the length of tape) 3426.

In a specific embodiment, the particle identification subsystem includesa pixel-to-tape mapping unit that maps a location of a particularparticle that has been captured within an image to the particle'sphysical location on the tape. The mapping unit determines a firstlocation of a particle within an image. The first location may be a setof pixel coordinates. For example, a pixel coordinate X may representthe particle's location as measured along an x-axis from a referencepoint in the image. A pixel coordinate Y may represent the particle'slocation as measured along a y-axis from the reference point in theimage. The pixel coordinates can be mapped into real space or into realx-y coordinates as measured from a reference point on the tape.

The particle collection cartridges may be assigned unique serial numbersso that images of the particles can be associated with correspondingcollection cartridge having the physical particles. As discussed, in anembodiment, the particle monitor includes a counter that tracks aposition of the tape. For example, the counter may track an amount orlength of tape taken up by the uptake reel, an amount or length of tapeunspooled from the supply reel, or both. Tracking the position of thetape allows for cross-referencing the images with the correspondingphysical particles on the tape.

In another specific embodiment, the tape may include a set of markersthat can be captured in the particle images. The markers may beindividually or sequentially numbered and distributed at variousintervals along a length of the tape. An interval may correspond to awidth of a field of view of the camera sensor so that a markerassociated with the interval will be captured in an image. The markerallows for cross-referencing the image with the portion of tape wherethe corresponding physical particles have been trapped. The markings maybe made using any technique for making a visible impression on the tapeincluding, for example, printing, silkscreen printing, stamping, orchemical processing. Alternatively, the tape may include a magnetizablelayer for magnetic marking and readout of tape locations.

At this point, status block 3440 contains a measurement status flag 3442and an analysis status flag 3444 with no bits set, and nullwork-in-progress and definitive particle classifications 3446 and 3448.This is the state of particle information packet 3400 at step 3505 ofFIG. 35.

At step 3510 (FIG. 35), the particle monitor embedded software storesinto data block 3480 (FIG. 34) sensor data 3481 for those RBG camerapixels including and surrounding the detected particle. At this step, abit in measurement status flag 3442 is set to indicate the capture ofcamera sensor data 3481. At this point, no decision has been madewhether the detected particle is even a biological particle rather than,for example, a dust particle.

The first analysis step is step 3515 (FIG. 35). This first analysis stepinvolves estimating the diameter or longest major axis of the detectedparticle. From this measurement there results a work-in-progress orpreliminary particle classification 3446 (FIG. 34) such as “<5 microns”(less than 5 microns), “10-15 microns” (between 10 and 15 microns),“35-40 microns” (between 35 and 40 microns) or “>200 microns” (greaterthan 200 microns). A corresponding bit in the analysis status flag isset. If the result had been outside the size of powdery mildew, a quickdefinitive particle classification 3448 of “not powdery mildew” wouldhave been made on the basis that the particle size is not compatiblewith the particles of interest 3434.

However, in this case we imagine a work-in-progress classification 3446that is compatible with the particles of interest. However, this sizerange may also compatible with many particles that are not of interest,such as dust particles that happen to be in this size range.

Given that the possibility remains that the packet might correspond to aparticle of interest, the software of the monitor makes a decision toanalyze particle shape. This is step 3520 of FIG. 35. If the outcome hadbeen a shape inconsistent with powdery mildew a definitive or finalparticle classification 3448 (FIG. 34) of “not powdery mildew” wouldhave been made.

However, we imagine a resulting work-in-progress classification 3446 ofmorphological characteristics (e.g., surface texture, shape, and size)consistent with powdery mildew. In engineering practice, thework-in-progress classification 3448 can be a numerical code that can beconfigured, by for example, scientists and software engineers or otherusers. As with all analysis steps, another bit in the analysis statusflag 3444 is set after completion of this analysis step.

Consider that the work-in-progress classification does not exclude thepossibility that the particle is powdery mildew. As a result, in step3525 (FIG. 35), the monitor's software makes a decision to analyze colorinformation in camera sensor data 3481 (FIG. 34). If the result had beena color not characteristic of powdery mildew, the processing of theparticle information packet 3400 would have ended with a “not powderymildew” definitive classification 3448. However, to provide a moreinstructive example, imagine step 3525 (FIG. 35) results in awork-in-progress classification 3446 (FIG. 34) of morphological andcolor characteristics consistent with powdery mildew.

Note that in steps 3520 (FIG. 35) and 3525, no new data is collected,only new analyses of previously measured data stored in data block 3480(FIG. 34).

At the completion of step 3525 (FIG. 35), the embedded software of themonitor has not yet reached a definitive classification, and may nothave the best information to decide what comes next. At such a point,the monitor looks for guidance from software on the cloud. In step 3530,the embedded software transmits the particle information packet 3400(FIG. 34) to the cloud.

The cloud software has access to great deal more information than doesthe embedded software of the particle monitor. For example, in step3535, the cloud software may have access to databases where the systemcollects and stores relevant information such weather patterns,elevations at various GPS coordinates, and historical records of powderymildew occurrences. Powdery mildew grows well in environments of highhumidity and moderate temperatures. The device GPS coordinates 3423 andtime stamp 3421 (FIG. 34) of particle identification block 3420 can beused to retrieve past or historical weather conditions. If thehistorical weather conditions indicate a lack of rain and humidity, thepresence of powdery mildew can be excluded. However, in FIG. 35 weconsider the case that the contextual information available on the clouddoes not exclude detection of particles of interest 3434 (FIG. 34).

To provide better discrimination between types of fungus, the cloudsoftware may decide that better morphology information is desirable andin step 3540 (FIG. 35) sends a request to the particle monitor tocollect further camera images of the particle with one or more alternatefocal depths. Focal depth may be varied by mechanical movement of lensand/or camera sensor, by electronic control of a variable lens, bysoftware control of processing of image data from a light-field camerasystem, or combinations of these.

The request received by the particle monitor triggers step 3545 and therequested measurements are made and added to the alternate focus data3482 (FIG. 34) of data block 3480. With this additional data, in step3550, a more refined shape or morphology analysis is performed. Theanalysis of step 3550 may be performed by embedded software of theparticle monitor (e.g., analyzed locally at the particle monitor), bycloud software (e.g., analyzed remotely by a cloud server), or both.

By capturing images of the particle at multiple focal depths, all partsof a particle can be brought into focus, thus providing more completemorphological information. For translucent particles, a scan of focaldepth may be used to capture three-dimensional particle structureinformation.

In step 3555, the cloud software also sends a request to the particlemonitor to collect further camera images with an alternate illuminationsource. In step 3560 the alternate illumination data is stored as item3483 (FIG. 34) in data block 3480.

Here we assume that the monitor is equipped with a quantum-dot LEDillumination source tuned to the absorption of powdery mildew. Thequantum-dot LED illumination source may be optional hardware that wasincluded in the particle monitor configuration due to the desire todeploy the monitor for agricultural monitoring; in this sense thedefinition of particles of interest 3434 may not only influenceprocessing of a particle information packet 3400, but also influence thehardware configuration of the particle monitor. Let us assume that sucha powdery mildew sensitive alternate illumination is used and in step3565 (FIG. 35), analysis (local, on the cloud, or both) of all the colordata in block 3480 (FIG. 34) confirms the presence of powdery mildew.

At this point, the evidence is strong that powdery mildew has beendetermined. However, before disturbing the agricultural managementconsultant with an alert, it may be prudent to obtain a second opinionfrom a human technician. In the scenario of FIG. 35, at step 3570, thecloud software sends a request for a second opinion from a trainedtechnician. In step 3575, resulting technician notes are stored in datablock item 3487 (FIG. 34). The request may include particle datacollected by the system and the system's determination based on theparticle data. This information may be displayed, for example, on anelectronic screen such as via a web page provided by the system. The webpage is accessible by the trained technician. The web page may furtherinclude an input box that the technician can use to enter notesregarding the accuracy of the identification and other information.

The technician may in turn request a third opinion by a scientificspecialist whose notes are captured in step 3580 and are stored in datablock item 3488 (FIG. 34). In this scenario, we imagine that thescientific specialist is fully convinced and the identification isdefinitively confirmed 3448 (FIG. 34). It is now time to inform theagricultural management consultant that powdery mildew has been detected(step 3585).

In the interests of cost and a fast response, it may well be moreexceptional than routine to involve humans, as in steps 3575 and 3580,in which it is otherwise an automated particle detection andclassification system. This exceptional scenario is considered here tomore fully describe a deeply multi-tiered particle discriminationscenario. A deeply multi-tiered scenario may also be one where the userof a particle monitoring device has required human review of theparticle data every so often (e.g., periodically). An example of this isan institutional, commercial, or single user with multiple systemsdeployed across a region and operating 24 hours a day 7 days a week. Insuch a scenario, human involvement as described in steps 3575 and 3580may take place as part of a quality assurance procedure.

For example, a human review of particle data may be required after onehundred, one thousand, ten thousand, a million, or another number ofparticle detections have occurred. A human review of particle data maybe required every hour, once a day, once a week, once a month or at someother interval of time. The criteria or frequency for when human reviewof particle data is required can be configurable such as by a user oradministrator of the system. The system (e.g., monitor, cloud server, orboth) can track this criteria to determine when a human review ofparticle data is required. When a human review of particle data isrequired, the system can send out a notification to the human reviewerto request a review of the particle data. The request may include theparticle data and an identification made based on the particle data. Thehuman reviewer can review the particle data to see whether or not theparticle identification was correct. Further leveraging of the talentsof the human reviewer may be provided by using results of human reviewsas input to automated machine learning algorithms so that reliance onhuman review decreases with time.

Even after the user has been alerted, the particle information packetmay continue to be processed for quality control and algorithmdevelopment purposes. In step 3590, the used reel of adhesive-coatedtape containing the detected particle is collected and stored in anarchive. Later, in step 3595, a bioassay using anti-bodies specific topowdery mildew is performed to verify beyond a shadow of a doubt thatthe particle was correctly classified—or to learn that a mistake wasmade and that the particle information packet should be closely studiedto determine what changes need to be made to the algorithms of thevarious tiers of the particle discrimination system. Depending on aparticle information packet's history, the data block 3480 may alsocontain alternate tape location camera sensor data 3484, informationfrom the cloud on local weather conditions 3485, information from thecloud on known seasonal pathogens 3486, laboratory microscope images3489 of archived particles, bio-assay data 3490, expert-technician notes3491 and/or expert scientist notes 3492. The particle information packetmay include a relationship block 3460 storing sequence numbers ofrelated particles 3462 and a nature of relationship of related particles3464.

Principles and aspects of the particle information packet may further beapplied to assessing the state of an identified particle such asassessing the state of the powdery mildew.

Principles and aspects of the system may also be applied to dronesconfigured for agricultural monitoring. One example of a drone is anunmanned aerial vehicle (UAV). Other examples of drones are unmanned,but not necessarily flying or aerial vehicle. Land-based drones mayinclude an autonomous 4-wheeler, sprayer, tractor trailer, and the likethat may be used in a cultivated field such as a vineyard. In anembodiment, a drone includes a particle monitor. The drone can beprogrammed to fly over an agricultural area (e.g., vineyard or farm) inorder to monitor for agricultural pathogens.

Table D below shows a flow for spore state discrimination according to aspecific embodiment.

TABLE D Step Description  1 Define a plurality of types of fluorescentbiomolecules of interest within the fungal spore.  2 Store a firstplurality of predetermined color characteristics of fluorescent lightfor each type of fluorescent biomolecules of interest, eachpredetermined color characteristic of the first plurality ofpredetermined color characteristics corresponding to excitement of arespective fluorescent biomolecule of interest under ultraviolet (UV)light having first spectral characteristics.  3 Store a second pluralityof predetermined color characteristics of fluorescent light for eachtype of fluorescent biomolecules of interest, each predetermined colorcharacteristic of the second plurality of predetermined colorcharacteristics corresponding to excitement of a respective fluorescentbiomolecule of interest under UV light having second spectralcharacteristics, different from the first spectral characteristics.  4Store a third plurality of predetermined color characteristics offluorescent light for each type of fluorescent biomolecules of interest,each predetermined color characteristic of the third plurality ofpredetermined color characteristics corresponding to excitement of arespective fluorescent biomolecule of interest under UV light havingthird spectral characteristics, different from the first and secondspectral characteristics.  5 Direct a flow of air comprising the fungalspore to a collection cartridge.  6 Trap the fungal spore within thecollection cartridge.  7 Illuminate the fungal spore in the collectioncartridge with visible light.  8 While the fungal spore is illuminatedwith the visible light, capture a first two-dimensional color image ofthe fungal spore.  9 Analyze the first two-dimensional color image toidentify an outline of the fungal spore, the outline of the fungal sporebeing defined by a set of image pixels receiving light from the fungalspore. 10 Illuminate the fungal spore in the collection cartridge withUV light of the first spectral characteristics. 11 While the fungalspore is illuminated with the UV light of the first spectralcharacteristics, capture a second color image of the fungal spore. 12Measure from the second color image a degree and color of fluorescencefor each pixel within the outline of the fungal spore. 13 Illuminate thefungal spore in the collection cartridge with ultra- violet (UV) lightof the second spectral characteristics. 14 While the fungal spore isilluminated with the UV light of the second spectral characteristics,capture a third color image of the fungal spore. 15 Measure from thethird color image a degree and color of fluorescence for each pixelwithin the outline of the fungal spore. 16 Illuminate the fungal sporein the collection cartridge with UV light of the third spectralcharacteristics. 17 While the fungal spore is illuminated with the UVlight of the third spectral characteristics, capture a fourth colorimage of the fungal spore. 18 Measurefrom the fourth color image adegree and color of fluorescence for each pixel within the outline ofthe fungal spore. 19 Based on the measurements from the second, third,and fourth color images of degree and color of fluorescence, estimate aconcentration of each type of fluorescent biomolecule of interest foreach image pixel within the outline of the fungal spore. 20 Generatingtwo-dimensional images of concentrations of the fluorescent biomoleculesof interest with outlines of fungal spores. 21 Based on information froma two-dimensional distribution of fluorescent biomolecules of interestwithin the outline of the fungal spore, determine the state of thefungal spore. 22 Correlate state of fungal spore with other variablessuch as time, temperature, humidity, and fungicide treatments. 23 Issuealerts and updates to responsible parties

In Step 1, the types of fluorescent molecules are of interest aredefined. In mathematical terms, this may include, for example, definingthe molecule enumeration index “j” of the equation of FIG. 30. In theexample in FIGS. 30 and 31, the molecule enumeration index “j” takes onthe values of “riboflavin”, “NADH” and “tryptophan”. This is areasonable choice for the determination of the state of biologicalparticles such as fungal spores or pollen grains. This is becauseflavins (see FIG. 46) and NADH (see FIG. 47) are fluorescingbiomolecules essential to cellular metabolism and tryptophan (see FIG.48) is an essential amino acid used in the construction of proteins.

If the particles of interest are diesel particulate matter and the stateof interest is the degree of chemical toxicity, then the defined typesof fluorescent molecules of interest may be polycyclic aromatichydrocarbons (PAHs) such as anthracene (see FIG. 41), triphenylene (seeFIG. 42), and coronene (see FIG. 43).

If particles of interest are diesel particulate matter, but pollengrains and mold spores are background particles that need to berejected, the molecules of interest may include riboflavin, NADH andtryptophan as well as PAHs.

It is desirable that for the given application that the set of moleculesof interest be ‘complete’ in the sense that no molecules outside the setcontribute significantly to measured fluorescent signals; this aidsinterpretation of the signals. More generally, in applying the describedsystems and techniques to a specific application, an important step isto determine (by experiment or literature research) the types offluorescent molecules of interest. Such a determination of the types offluorescing molecules of interest may well be done by engineers orscientists associated with the company providing the particle-monitoringdevice 700. The resulting information may be coded into software orinitialization files associated with the device. For example, theinitialization files may be stored in the particle monitor device. Theinitialization files may be transmitted from a central server, over anetwork, and to the particle monitor device. The initialization filescan be updated remotely, locally (e.g., copied from pluggable USB driveinserted into monitor device), or both. In embodiment, a method includesstoring first initialization at a particle monitor device where thefirst initialization files specify first fluorescent molecules ofinterest; receiving at the particle monitor second initialization fileswhere the second initialization files specify second fluorescentmolecules of interest, different from the first fluorescent molecules ofinterest; and replacing the first initialization files with the secondinitialization files.

Taking the example of riboflavin, NADH and tryptophan as the types offluorescent molecules of interest, in Step 2 the fluorescent behavior ofthese molecules of interest in response to a particular UV illuminationsource is determined and stored for later use in Step 19. For example,particle-monitoring device 700 may include a UV illumination source witha representative wavelength of 365 nm that results in riboflavinfluorescing with a green color having a representative emissionwavelength of 520 nm, NADH fluorescing with a blue color having arepresentative emission wavelength of 455 nm, and tryptophan notfluorescing at all. It is to be understand that a 365 nm UV lightsource, such as a 365 nm UV LED, does not only generate photons ofwavelength 365 nm, but rather generates photons with a wavelengthspectrum of finite width and peaking around 365 nm. It is the colorcharacteristics of emitted fluorescent light in response to illuminationby the spectrum from the UV light source that is of interest. It is tobe understood that in concisely referring to an excitation wavelength of365 nm from a UV light source, we mean the entire spectrum from the UVlight source. Similarly in concisely referring to a fluorescent emissionwavelength of, say, 520 nm, it is understood that a full description ofthe color of fluorescently emitted light is a spectrum peaking near 520nm. In FIGS. 30 and 31, the color characteristics of the fluorescentmolecules riboflavin, NADH and tryptophan under 365 UV illumination arenotated with the letter “F” with subscript “RIBOFLAVIN” and superscript“365 nm→520 nm”, the letter “F” with subscript “NADH” and superscript“365 nm→455 nm”, and the absence of a letter “F” with subscript“TRYPTOPHAN” with superscript “365 nm→ . . . ”. The quantitative values,or absences, of these “F” values quantify a plurality of predeterminedcolor characteristics of fluorescent light for each type of fluorescentmolecule of interest in response to one particular type of UV lightsource (e.g. a 365 nm UV LED) having its own UV spectralcharacteristics. Determination of these “F” values, or similarinformation in a different format, may be done by engineers orscientists associated with the provider of particle-monitoring device700, and then coded into software or initialization files associatedwith the device.

In some applications, one UV illumination source may be sufficient. Inother applications it may be desirable to enrich measured data with theaid of a second UV light source with different spectral characteristics.For example, particle-monitoring device 700 may include a second UV LEDwith a representative wavelength of 325 nm. If so, the method proceedsto Step 3 that repeats the previous step for the second UV light source.In the example of FIGS. 30 and 31, Step 3 may result in thedetermination of “F” values for 325 nm→520 nm riboflavin fluorescenceand 325 nm→455 nm NADH fluorescence, and an absence or zero value of an“F” value for tryptophan.

If particle-monitoring device 700 includes a third UV light source, thenoptional Step 4 applies. For example, particle-monitoring device 700 mayinclude a third UV LED with a representative wavelength of 280 nm. Inthe example of FIGS. 30 and 31, Step 4 may result in the determinationof “F” values for 280 nm→520 nm riboflavin fluorescence and 280 nm→400nm tryptophan fluorescence, and an absence or zero value of an “F” valuefor NADH. While not included in Table D, further steps may be insertedto accommodate additional UV light sources. Each additional UV lightsource enriches the data available but also adds cost. In manyapplications the best balance between cost and performance may be withone, two or three UV light sources.

As seen in FIGS. 28, 29 and 31, as well as the earlier discussion ofthese figures, riboflavin fluorescence is also possible with 445 nm bluelight. It is to be understood that in Steps 2, 3 or 4 above, ‘UV light’may be interpreted as excitation light of sufficiently short wavelengthto induce visible-light fluorescence of molecules of interest, which insome cases the excitation light may be within the visible lightspectrum.

In the example of FIG. 28S, each molecule of interest has only oneemission wavelength peak value independent of the excitation wavelength.For example, riboflavin emits a spectrum of wavelengths with a peakwavelength about 520 nm, independent of whether the excitationwavelength is 445 nm, 365 nm, 325 nm or 280 nm. This behavior istypical. However, if a molecule had two fluorescence emission peaks,that could easily be accommodated with additional “F” values.

In Steps 5 and 6, particles of interest are captured within thecollection cartridge and moved to the inspection zone as describedpreviously. A visible light image is captured in Steps 7 and 8. A firstfluorescence image under UV light illumination is captured in Steps 10and 11. Optional second and third fluorescence images under second andthird UV light sources are captured in steps 12 and 13 and steps 15 and16 respectively. While the visible light image is captured first inTable D, the images may be captured in any order. Furthermore, thecapture of additional UV fluorescence images of Steps 12 and 13, and ofSteps 15 and 16, may be conditional on whether the previously capturedimages are deemed sufficient to determine the state of the particles ofinterest. For all images, two-dimensional color information is capturedin the form of RGB pixel values from the camera sensor 1420. Such colorimage data is represented by “RGB(x,y)” with a subscript of either“RED”, “GREEN” or “BLUE” in FIGS. 30 and 31 where each pixel hasdifferent coordinates “(x,y)”. RGB color pixel data for UV-excitedfluorescence is captured much the same way as for visible lightscattering, but the captured image data is processed differently.

In Step 9, the visible light image from Steps 7 and 8 is analyzed todetermine outlines of particles of interest, that is, it is determinedwhich camera sensor pixels correspond to (x,y) locations insideparticles of interest and which to (x,y) outside particles of interest.Alternatively, one or more of the UV fluorescence images, perhaps incombination with the visible light image, is used to determine particleoutlines. After completion of Step 9, UV fluorescence color image datacorresponding to pixels within particles of interest may be determinedas in Steps 12, 15 and 18.

The steps leading up to Step 19 provide rich image data not only torecognize particles of interest, but also to determine their state. Inparticular, in Step 19, the concentration of each type of fluorescencemolecule of interest may be determined from the captured image data, themolecule fluorescence color characteristics determined in Steps 2, 3 and4 as well as the pixel color sensitivities of camera sensor 1420. InFIGS. 30 and 31, the pixel color sensitivities are represented by theletter “p” with a subscript specifying the pixel color (“RED”, “GREEN”or “BLUE”) and an argument specifying the wavelength of thefluorescently emitted light, or more generally the wavelength spectrumof the fluorescently emitted light. The numerical “p” values may bedetermined along with “F” values in Steps 2, 3 and/or 4. Referring tothe formula of FIG. 30, the RGB(x,y) values within particles of interestfor the one or more UV fluorescence images are known from Steps 12, 15and/or 18. The “p” and “F” values are known from Steps 2, 3 and/or 4.Using this information and solving the resulting set of simultaneouslinear equations such as illustrated in FIGS. 30 and 31, theconcentrations “C(x,y)” for each molecule of interest may be determinedas a function of pixel location (x,y). For example, if the molecules ofinterest are riboflavin, NADH and tryptophan, the resultingconcentration maps may be notated as “C_(RIBOFLAVIN)(x,y)”,“C_(NADH)(x,y)” and “C_(TRYPTOPHAN)(x,y)”.

Steps 7 and 8 may be combined with Steps 10 and 11. That is, during anexposure for a camera sensor image capture, the particles of interestmay be illuminated by visible light (Step 7) and by UV light (Step 10),either simultaneously or sequentially. Because exposure times for UVlight are typically longer than for visible light, the visible lightsource might flash for a fraction of a second during UV illuminationlasting several seconds. A shorter visible light illumination may occurbefore, during or after UV illumination; if during the visible lightillumination may occur at any time during the UV illumination period.The resulting captured image serves as both the scattered visible lightimage of Step 8 and the fluorescence image of Step 11. From this singlecombined RGB image, the fungal spore outline may be determined (Step 9)and the fungal spore fluorescence color may be determined (Step 12).Likewise Steps 7, 8 and 9 may be repeated and combined with Steps 13, 14and 15 as well as with Steps 16, 17 and 18.

In a specific embodiment there are four illumination sources that inplan view illuminate the inspection zone 840 from the four compassdirections. Illumination directions may be at an angle of 60 degreeswith respect to the vertical optical axis of the camera sensor. Thenorth illumination source is a UV LED while the east, south and westillumination sources are white LEDs. In an embodiment all LEDs generateun-polarized light. The camera sensor exposure time is 15 seconds duringwhich the UV LED is on. After 14 seconds of exposure and UVillumination, the east white LED is turned on for 0.033 seconds, afterwhich the south LED is turned on for 0.033 seconds and then the west LEDis turned on for 0.033 seconds. For at least some types of mold spores,it has been observed that images captured under such illuminationconditions provide excellent data for extracting morphological dataincluding a determination of the outlines of spores. In someapplications it may be sufficient to drop the south white LED have onlytwo white LEDs illuminating from east and west directions. More thanthree white LEDs may be desired in other applications. Other methods ofenabling white-light illumination from multiple directions are alsopossible. This includes providing a light ring so that illuminationcomes from all directions (in plane view). This also includes providingmechanical means to physically move a white LED so that it canilluminate the inspection zone from different directions, e.g. bymounting LED on a turret.

In an embodiment, the calculation of molecule concentrations C(x,y) willbe most reliable if the set of molecules of interest from Step 1 iscomplete (in the sense that no other molecules contribute significantlyto the measured fluorescence signals). It is also desirable that thenumber of equations corresponding to FIG. 30 exceeds the number ofmolecules of interest to that the simultaneous linear equations areover-constrained; a poor quality of fit would provide a warning that theset of molecules of interest is not complete.

An engineer or scientist may find it intuitive and informative toinspect the molecule concentration maps in a visual format. For example,software might display an image of a particle of interest where thecomputer display (not camera sensor) pixel at location (x,y) has a redsub-pixel value set to the value of C_(RIBOFLAVIN)(x,y), a greensub-pixel value set to the value of C_(NADH)(x,y), and a blue sub-pixelvalue set to the value of C_(TRYPTOPHAN)(x,y). This corresponds to Step20. From the information of Step 20, the state of the particles ofinterest is determined by a skilled human or by automatic software inStep 21.

If artificial intelligence (AI) or neural network algorithms are trainedand applied, such machine-learning methods may well directly process theRBG(x,y) image data, thus skipping Steps 19 and 20 and proceedingdirectly to the particle state determination of Step 21. In other words,in a machine-learning trained-neural-network calculation, theinformation of Steps 19 and 20 may be present but deeply hidden withinthe weights of a neural network calculation.

In a specific embodiment, a reference fungal spore in a known state isilluminated using UV light at various spectral characteristics (e.g.,different UV wavelengths). The fluorescence from the biomolecules inthis reference fungal spore with the known state is captured andrecorded as predetermined color characteristics of fluorescent light.These predetermined color characteristics of fluorescent light thenfunction as references to identify a state of a fungal spore at issue.

In a specific embodiment, the fungal spore at issue is illuminated withUV light of a particular spectral characteristic. An image is captured.Concentrations of biomolecules of interest (e.g., riboflavin, NADH, andtryptophan) are estimated from the image. These estimated concentrations(generated from the image taken while the fungal spore at issue wasilluminated with UV light of the particular spectral characteristic) arethen compared against a subset of the predetermined colorcharacteristics of fluorescent light. Each predetermined colorcharacteristic of fluorescent light in the subset corresponds to UVlight of the same particular spectral characteristic under which theimage was taken. Upon finding a match between the estimatedconcentrations and one of the predetermined color characteristics offluorescent light from the subset, the matching predetermined colorcharacteristic may be cross-referenced to its reference fungal spore ofthe known state. In embodiment, the predetermined color characteristicsof fluorescent light for fluorescent biomolecules of interestcorresponds to a fungal spore of a known state having a particularconcentration of those fluorescent biomolecules of interest.

In Step 22, the latest particle state information from Step 21 iscombined with previous particle state information as well as withinformation on other variables such as time, temperature, humidity, andfungicide treatments. For example, if when temperature and humidity isfavorable to mold infestations and the number of spores detected in avirulent state is increasing, it may be concluded that a moldinfestation is imminent. As another example, if after a fungicidetreatment, the number of detected spores in a healthy state rapidlydecreases to zero, then there is reason to believe that the fungicidetreatment was successful and sufficient. On the other hand if afungicide treatment does not reduce the number of healthy spores, it maybe an indication that the mold has developed a resistance to thefungicide in use and a different fungicide is needed. The analysis ofdata in Step 22 may lead, in Step 23, to alerts or updates being issuedto responsible parties. For example, if Step 22 gives reason to believethat an outbreak of mold infestation is imminent, a message to theresponsible farmer may be sent recommending an immediate application offungicide.

FIGS. 36-37 show the placement of a particle monitor 3605 (FIG. 36) in avineyard. In this specific embodiment, the particle monitor is poweredby a rechargeable lead-acid battery 3705 (FIG. 37) which in turn isrecharged by a set of solar panels 3710.

To prove operability, a prototype of the particle monitor was fullybuilt, deployed within a vineyard, tested, and verified to operate asintended. As shown in FIG. 36, in this specific embodiment, the particlemonitor includes an eyelet 3610 connected to a top end of the particlemonitor and a set of wires 3615 extending from a bottom end of theparticle monitor.

The eyelet allows a hook or carabiner 3620 to pass through so that theparticle monitor can be attached to a vineyard wire 3625 and suspendedin mid-air. Locating the particle monitor above the ground surface helpsto facilitate a good sampling of air and helps to prevent ground dirt,dust, and bugs (e.g., ants) from entering the particle monitor.

An opposite end of the wires is connected to the rechargeable lead-acidbattery (FIG. 37) which, as discussed, is recharged by the solar panels.In this specific embodiment, the battery and solar panels are locatedexternal to or outside the particle monitor. Locating the batteryexternal to the particle monitor helps to reduce the overall weight ofthe particle monitor. For example, as shown in FIG. 36, the particlemonitor can be hung from an existing vineyard wire without the vineyardwire breaking or experiencing heavy sagging due to the weight of theparticle monitor. Locating the battery external to the particle monitoralso helps to reduce the need for a lightweight and long-lastingbattery; such characteristics generally increase the cost of thebattery. The battery can be a relatively large lead-acid battery.Lead-acid batteries are generally less expensive than other batterytypes such as nickel cadmium (NiCd), nickel-metal hydride (NiMH),lithium ion (Li⁺ ion), and lithium ion polymer (Li⁺ ion polymer).Multiple particle monitors (e.g., two or more particle monitors) may bepowered by the same external battery.

Locating the solar panels external to the particle monitor allows thesolar panels to be placed in a location away from shading so that lotsof sunlight can be received. The particle monitor can be placedproximate to or near a grape vine (as shown in FIG. 36). The shading ofthe particle monitor by the leaves of the grape vine will notdetrimentally affect the powering of the particle monitor because thesolar panels are located external to the particle monitor.

Diesel Exhaust Monitoring

The above discussion described detection of spores of agriculturalinterest. Nevertheless, the presented techniques are more broadlyapplicable. A noteworthy example of an alternate application of theabove methods is the detection of soot particles in the air, such asfrom the exhaust of diesel engines. Due to the impact of diesel exhaustparticles on human health, monitoring of soot particles is of particularinterest to the field of air pollution monitoring.

Soot particles are formed by incomplete combustion of hydrocarbon fuels.For example, consider combustion of paraffin in a candle flame. Paraffinis composed of hydrocarbons with molecular formulas C_(N)H_(2N+2) suchas C₃₁H₆₄. Taking this N=31 example, complete burning with oxygen fromthe air results in only carbon dioxide and water vapor via the chemicalreaction C₃₁H₆₄+47 O₂→31 CO₂+32 H₂O. If a metal spoon is placed wellabove the candle flame, it remains clean. When the metal spoon is moveddown into the top of the candle flame, cooling the flame's gas andhalting the chemical reactions before combustion is complete, a blacklayer of soot is formed on the spoon's surface. Incomplete burning ofdiesel fuel and other hydrocarbon fuels also generates soot.

The black color of soot correctly suggests that soot particles arecarbon rich. Note that for complete combustion, such as C₃₁H₆₄+47 O₂→31CO₂+32 H₂O, the hydrogen atoms of the hydrocarbon fuel are completelyseparated from the carbon atoms. During incomplete burning, chemicalreactions occur that only partially separate hydrogen and carbon atoms,and only partially combines carbon atoms with oxygen to form carbondioxide. This results in carbon rich molecules with formulas of the formC_(N)H_(M) where M<2N+2. Production of molecules including aromaticrings is favored due to their relative stability. The simplest aromatichydrocarbon is benzene whose chemical formula is C₆H₆ which is indeedcarbon rich with 6=M=N<2N+2=14.

FIGS. 38-40 illustrate three representations of the planar benzenemolecule C₆H₆. With heat and limited oxygen supply, aromatic rings maycombine to produce polycyclic aromatic hydrocarbons (PAHs), namelyhydrocarbons whose molecules contain multiple aromatic rings. Examplesof polycyclic aromatic hydrocarbons (PAHs) are anthracene, triphenyleneand coronene with chemical formulas C₁₄H₁₀, C₁₈H₁₂, and C₂₄H₁₂respectively. FIG. 41 shows an anthracene molecule. FIG. 42 shows atriphenylene molecule. FIG. 43 shows a coronene molecule.

Soot particles contain polycyclic aromatic hydrocarbons (PAHs) ofvarying degrees of complexity. Many polycyclic aromatic hydrocarbon(PAHs) molecules are toxic. The nature and amount of polycyclic aromatichydrocarbons (PAHs) in soot particles in the air we breathe is ofinterest from the perspective of human health.

An important characteristic of airborne soot particles is their size.The soot formed on a metal spoon in a candle flame appears to the humaneye to be a continuous and smooth black coating, but in fact is composedof many microscopic soot particles. Diesel exhaust also contains suchmicroscope soot particles. The largest airborne soot particles(including particles that are aggregates of smaller particles) can be aslarge as several microns in diameter.

However, for diesel particulate matter (DPM), the vast majority ofairborne soot particles are of sub-micron size including the100-nanometer sizes and below. Unlike larger particles, sub-micron sizedsoot particles travel deep into our lungs and for this reason are ofparticular concern to the Center for Disease Control's (CDC's) NationalInstitute of Occupational Safety and Health (NIOSH). NIOSH method 5040for monitoring airborne diesel particular matter (PDM), which isincorporated by reference, is designed to detect sub-micron sizedparticles. For example, the NIOSH 5040 document, available at the CDCwebsite and incorporated by reference, includes the following quoteconcerning the removal of micron-sized particles from the particulatesto be measured: “ . . . For measurement of diesel-source EC in coalmines, a cyclone and impactor with submicrometer cutpoint are requiredto minimize collection of coal dust. A cyclone and/or impactor may benecessary in other workplaces . . . ”. The sub-micron sizes of dieselexhaust soot particles of interest are much smaller than themultiple-micron sizes of mold spores and pollen grains.

Given the small sizes of airborne soot particles of interest, it wouldat first appear that a microscope-based device would be of little valuein the field of diesel particulate matter (PDM) monitoring. The size ofa 100-nanometer soot particle is much smaller than the wavelengths ofvisible light. (Visible light wavelengths range from 400 nanometers atthe violet to 700 nanometers for red.) Optical microscopes cannot imageobjects that are small compared to the wavelength of light.Morphological features of such small soot particles are far beyond theresolution of optical microscopes. Electron microscopes can image sootparticles with high resolution, but with high cost and complex samplepreparation are not amenable to low-cost real-time monitoring. Smallsoot particles of interest are difficult to detect at all within anoptical image, even as a small blurry dot. Per pessimistic conventionalthinking, optical microscope-based devices have little to offerregarding the needs of diesel particulate matter (DPM) monitoring.

Applicant has identified a solution that goes contrary to theconventional thinking. Applicant has observed that the NIOSH 5040 methodfor diesel particulate matter (DPM) monitoring is far from perfect. Itmeasures the total mass per unit air volume of ultrafine dieselparticulate matter (DPM), but provides no information on the particlesize distribution. Two air samples with exactly the same amount ofdiesel particular matter (DPM) in units of micrograms per liter, butwith different particle size distributions, may well have very differentdegrees of impact on human health.

Furthermore, even with the same micrograms per liter and the sameparticle size distributions, two air samples may still have differentimpacts on human health if their chemical compositions differ. Forexample, soot particles in one air sample may contain more toxicpolycyclic aromatic hydrocarbons (PAHs) than the other. There is a needto complement the micrograms-per-liter measurement of airborne dieselparticulate matter (DPM), such as provided by NIOSH 5040 methods, withan ability to monitor other characteristics of diesel particulate matter(PDM) such as particle size distribution and polycyclic aromatichydrocarbon (PAH) content.

Applicant has realized that an examination of the tail of particle sizedistribution can be useful in diesel exhaust monitoring. For sootparticles that are large enough to be imaged by an optical microscope,optical images provide a good means via color and morphology todifferentiate between larger soot particles and other particles ofsimilar size. Hence the upper end of the soot particle distribution canbe measured even in the presence of background particles of similarsizes. This is not true for monitors based on the NIOSH 5040 method.However, as discussed above, it is understood that such larger sootparticles that can be effectively imaged in an optical microscope arenot in the size range of greatest interest to human health.Nevertheless, surprisingly, and as described below, monitoring of largesoot particles does nevertheless provide information of interest.

The plot in FIG. 44 illustrates two size distributions of dieselparticulate matter (DPM). Distribution A has on average smallerparticles sizes and a larger total number of particles whiledistribution B has on average larger particle sizes and a smaller totalnumber of particles. Consider the case where both distributionscorrespond to exactly the same mass per liter of air as measured by theNIOSH 5040 method.

Distribution A will be more damaging to human health as it contains moreparticles making it deep into our lungs and provides a larger totalparticulate surface area for exposing lung tissue to toxic polycyclicaromatic hydrocarbons (PAHs). Now consider the larger-size tails of thetwo distributions, such as the large-size tails above 1 micron in size.Distribution B has a much bigger tail into micron sizes thandistribution A, something that can be observed with anoptical-microscope based monitoring device. By measuring dieselparticulate matter (DPM) with both NIOSH-5040 methodology andsimultaneously with optical-microscope based devices, usefulparticulate-size-distribution information is provided that is relevantto assessing the effects of the diesel particulate matter (DPM) on humanhealth.

Larger particles settle out of air faster than smaller particles. For agiven source of diesel particulate matter (DPM), the distribution ofparticle sizes will change over time. For example, on a calm day, themicron-sized particles will settle out of the air faster than thesub-micron-sized particles. This is illustrated FIG. 45 in which themicron sized particles in the source distribution C is missing from thesize distribution D of the polluted air after it has separated from thesource in time and distance. The presence or lack of theabove-one-micron tail of the size distribution reveals clues about thedistance of the source from the monitoring location. Theabove-one-micron tail of the size distribution, even if empty, providesuseful additional information regarding diesel particulate matter (DPM)that is not available from NIOSH-5040 method measurements alone.Depending on the application, the optimal choice of threshold size forthe large-size distribution tail may vary from a value of one micron,but the principles remain the same.

More generally, monitoring of the above-one-micron tail of the dieselparticulate matter (DPM) distribution provide additional independentinformation that may be used in various ways by resourceful airpollution scientists and technicians. Perhaps an appropriate analogyhere is between the tip of an iceberg and the above-one-micron tail ofdiesel particulate matter (DPM), and between sub-micron distribution ofdiesel particulate matter (DPM) and the undersea portions of icebergs.It is the undersea portions of icebergs that sink ships. Nevertheless itis useful for sailors to look for tips of icebergs rising above thesurface of the sea. Likewise it is the sub-micron distribution of dieselparticulate matter (DPM) that is most damaging to human health, but itmay nevertheless useful to monitor the above-one-micron tail of dieselparticulate matter (DPM) distributions.

Probing PAH Toxicity with UV Fluorescence

Under UV illumination, some polycyclic aromatic hydrocarbons (PAHs)fluoresce. For example, this is true for the above-mentioned anthracene(C₁₄H₁₀), triphenylene (C₁₈H₁₂) and coronene (C₂₄H₁₂). For example,anthracene fluoresces blue under UV light. Thus soot particles mayinclude UV fluorescent molecules analogous to the flavin, NADH andtryptophan molecules of biological particles discussed above. Themolecular structures of flavin, NADH and tryptophan are shown FIGS. 46,47, and 48, respectively.

Like the polycyclic aromatic hydrocarbons (PAHs) anthracene,triphenylene and coronene, these three biological molecules fluorescevisible light under UV illumination due to the presence of polycyclicaromatic rings. The techniques described above in connection with FIGS.29, 30 and 31 can equally well be applied to the analysis of polycyclicaromatic hydrocarbons (PAHs) content of soot particles. Data related tothe polycyclic aromatic hydrocarbon (PAH) content of soot particles maybe relevant to the assessment of the chemical toxicity of dieselparticulate matter (DPM).

The polycyclic aromatic rings of the three biological molecules in FIGS.46-48 differ chemically from the polycyclic aromatic rings of polycyclicaromatic hydrocarbons (PAHs). In the latter case, the rings contain onlycarbon atoms, while for the biological molecules rings also includenitrogen atoms. Given this chemical difference between fluorescingmolecules in soot particles and biological particles (like spores andpollen), the two different types of particles are expected to havedistinct UV fluorescence properties. These differences can be exploitedby taking advantage of the three color images of RGB camera sensors aswell as the option of the devices described above to use UV illuminationof one or more wavelengths.

During the course of field studies, it was observed that dusting a cropwith fungicide dusting resulted in detected particles having strongfluorescence under 340 nm UV illumination as well as having sufficientsizes to be imaged by optical microscopy. In light of the abovediscussion, and that fact that many fungicides include aromatic ringswithin their molecular structures, similar observations may be fordustings or sprays of a number of fungicides. Examples of fungicideswith aromatic rings of various sorts include Metalaxyl, Bupirimate,Carendazim, Boscalid, Azoxystrobin, Cyprodinil, Proquinazid, Quinoxyfen,Iprodione, Spiroxamine, Fenhexamid, Dimehtomorph, Chlorothalonil andMetrafenone. More generally, a number pesticides (“pesticides” includeherbicides and insecticides as well as fungicides) may well have strongUV fluorescence signatures.

UV Fluorescence of Sub-Micron Soot Particles

Distinct color characteristics of soot particle UV fluorescence mayenable detecting sub-micron soot particles that would otherwise beundetected by an optical-microscope based system. Consider, as anexample, that the adhesive surface of the tape described above collectssub-micron soot particles in addition to biological particles likepollen and spores as well as inorganic dust. It may well be impossibleor difficult to confidently recognize sub-micron soot particles in RGBcamera sensor images under white light illumination.

However, a general fluorescent glow of the adhesive coated tape under UVillumination may provide a signature and measure of a dusting ofsub-micron soot particles. In particular, a fluorescent glow might beidentified as having the fluorescent color characteristics of the sootparticles' polycyclic aromatic hydrocarbons (PAHs). The fact thatinorganic dust does not fluoresce eliminates it as a potentialbackground. The ability of the above-described devices to image andrecognize biological particles is also an advantage here. Anycamera-sensor pixels within images of pollen, spores and otherbiological particles can be removed before analysis of the color of anyremaining fluorescent glow; this is a powerful way to minimize UVfluorescence background from biological particles.

As just noted, an ability to recognize pollen, spores and otherbiological particles within RBG camera sensor images enables powerfulbackground-subtraction methods when seeking optical signals from anothersource of interest such as diesel particulate matter. Both morphologyand color, including UV fluorescence excitation and emission wavelengthcharacteristics, provide techniques to recognize pollen, spores andother biological particles.

The simplest morphological feature is size. Pollen and spores tend to bemany microns while most diesel particulate matters are submicron insize. That is, pollen and spores tend to extend over many pixels in RGBcamera sensor images while diesel particles generally do not.Furthermore, pollen grains or mold spores are typically large enough toreveal distinctive shapes in optical images.

Color is another differentiator. Soot particles tend to be opaque andblack while pollen and spores tend to be more translucent. In general,any image processing and image recognition techniques that maybe used toidentify pollen, spores and other biological particles when they areobjects of interest may also be used to identify pollen, spores andother biological particles when they are backgrounds to be removed fromanalysis while pursuing other optical signals of interest.

As an analogy here, consider a stargazer looking at light from the MilkyWay. The human eyes and mind are able to see past bright foregroundstars and see the dim Milky Way behind it. However, if one attempted todetect the Milky Way by pointing a light meter in the general directionof the Milky Way, the dim light from the Milky Way would be lost in thebackground of the bright foreground stars. Likewise, UV fluorescencedetection without camera-sensor imaging may leave an unusable sub-micronsoot particle signal that is swamped by biological particlefluorescence. However, like a stargazer's eyes and mind can ignoreforeground starts, the particle monitors described above have theability to separate bright fluorescence from biological particles from apossibly much dimmer fluorescence signal from many small soot particlescaptured on the adhesive surface.

Combining Scanit & NIOSH 5040 Detectors

The methods and apparatuses described above may be combined with otherparticle detection methods such as those described in the NIOSH 5040method for airborne diesel soot detection. A NIOSH 5040 method baseddetector may include a transparent or translucent air filter throughwhich sampled air passes after larger particles have been removed. Assoot collects on such an air filter, it darkens and becomes lesstransparent or translucent. A light source may be placed on one side ofthe air filter and a light detector placed on the other side of the airfilter. A reduction in light transmission through the air filterprovides a technique to estimate the micrograms of diesel soot per literof sampled air. Such a methodology may be combined with the methods andapparatuses described above.

For example, a particle-monitoring device 5100 of FIG. 51 is such amodification of the particle-monitoring device 700 shown in the samevertical cross-sectional view as FIG. 13. Blower 5110 drives airflowthrough the particle-monitoring device 5100 as indicated by arrows 1320,1330, 5130, 5132, 5134, and 1340.

Before reaching gap 1317, this airflow is intercepted by transparent ortranslucent air filter 4910. Translucent air filter 4910 is asufficiently fine filter to capture any diesel particulate matter in theair. This translucent air filter 4910 is mounted within a removableair-filter cartridge 4905. (For clarity, sliding doors and othermechanics for inserting or removing air-filter cartridge 4905 are notshown. Also for clarity, not shown are various gaskets and other airflowbarriers needed to avoid undesired airflow paths.)

In some embodiments, the air-filter cartridge 4905 may be removed andshipped to a lab for analysis while a replacement fresh cartridge may beinserted into the particle-monitoring device 5100.

In other embodiments, the accumulation of any diesel particulate matteron air filter 4910 is optically monitored in situ. For this purpose,particle-monitoring device 5100 may be optionally provided with filterlight source 5010 and filter light detector 5020. For clarity, mountingand electrical connections of the filter light source 5010 and thefilter light detector 5020 to the particle-monitoring device 5100 arenot shown.

Filter light source 5010 may be a visible light LED and filter lightdetector may be a phototransistor. Alternatively, filter light source5010 and/or filter light detector 5020 may be ends of optical fibersthat transmit light to or from electro-optical components on a circuitboard elsewhere within particle-monitoring device 5100. Reduction of thesignal from the filter light detector 5020 provides a measure of thedarkening of the translucent air filter 4910 due to accumulation ofdiesel particulate matter or other sooty material in the air. Such insitu optical measurements may be in addition to, or instead of,laboratory analysis of accumulated diesel particulate matter or othertypes of soot.

In yet other embodiments, air-filter cartridge 4905 and particle-mediacartridge 805 are designed to be sufficiently similar in mechanicaldesign and form factor so that particle-media cartridge 805 can beremoved from its location shown in FIG. 51 and replaced with air-filtercartridge 4905, and it so doing placing translucent air filter 4910within the field of view of the optical system of RGB camera sensor1420. This enables two-dimensional image analysis of matter accumulatedin the air filter 4910. If opaque objects unrelated to dieselparticulate matter are observed in images from RGB camera sensor 1420,then appropriate corrections may be made to previous opticaltransmission measurements from filter light source 5010 to filter lightdetector 5020.

If the illumination system associated used with RGB camera sensor 1420includes UV light sources, the fluorescence properties of materialcollected by the air filter 4910 may be measured. This is of interest asa probe of the presence of toxic compounds within any accumulated dieselparticulate matter. For example, UV fluorescence properties ofaccumulated diesel particulate matter may provide a technique to detecttoxic compounds such as anthracene, triphenylene, coronene, and others.

FIG. 49 illustrates an air-filter cartridge 4905 designed for insertioninto the slot that normally holds a particle-media cartridge 805 as wellas for insertion into the air-filter cartridge 4905 location shown inFIG. 51. FIG. 49 shows a cross-sectional view similar to that of FIG. 9.Air filter support 4920 contains a hole 4925 through which air may flowafter passing through translucent air filter 4910. The translucent airfilter 4910 is held in place by a retaining structure 4930.

Retaining structure 4930 may be of any design that services its purpose,including a plastic piece held in place by a tight friction fit, or aslightly concave piece of thin sheet metal. Items 4950, 4960 and 4970are explained with the aid of FIG. 50.

FIG. 50 is a modification of FIG. 49 with additions illustrating airflowand optical components. When inserted into particle-monitoring device5100 as shown in FIG. 51, air flows through air-filter cartridge 4905.Air leaving blower 5110 and approaching air-filter cartridge 4905 isindicated by arrow 5050. Air that flows through air-filter cartridge4905, as illustrated by arrows 5052 and 5054, leaves the cartridgethrough exit hole 4970, as indicated by arrow 5056.

If it is desired to shield optical components from background lightentering through exit hole 4970, an optical baffle 4960 may be included.The optical-detector hole 4950 of air-filter cartridge 4905 is similarin geometry to gear-shaft hole 850 of particle-media cartridge 805 (seeFIG. 8). Optical-detector hole 4950 avoids mechanical interference withthe gear shaft when air-filter cartridge 4905 is inserted into the slotnormally occupied by particle-media cartridge 805. The optical-detectorhole 4950 serves a second purpose. It allows filter light detector 5020to be located below air filter 4910 within the volume of the air-filtercartridge 4905, and yet be mechanically attached and electricallyconnected to the particle-monitoring device 5100, and not mechanicallyattached to or mechanically interfering with the body of air-filtercartridge 4905.

Filter light source 5010 is also mechanically attached and electricallyconnected to the particle-monitoring device 5100. Dashed arrow 5030illustrates a light path from filter light source 5010 through airfilter 4910 to the filter light detector 5020. Any diesel particulatematter within air flowing along arrow 5050 will accumulate on air filter4910 making it darker and reducing the light intensity reaching filterlight detector 5020. In this manner measurements based on the NIOSH 5040method may be performed in real time.

Returning to FIG. 51, optionally, particle-monitoring device 5100 mayinclude a course air filter 5140 to reduce the rate that largerparticles, such as those not related to diesel particular matter, reachair filter 4910. Instead, or in addition, a cyclone impactor or othertype of impactor (not shown) may be inserted in the airflow upstream ofthe air filter 4910 to further prevent any larger particles reaching airfilter 4910.

The above discussion in connection with FIGS. 49, 50 and 51 demonstratethat it is possible to combine RGB camera sensor based measurementmethods and NIOSH 5040 methods in a way that combines their respectivestrengths.

Fungicide (Pesticide) Particle Detection

Returning to agricultural applications, it is of interest to be able todetect and recognize particles of pesticide sprays or dustings. As notedabove, such pesticide particles may well be sufficiently large to beimaged in RGB camera sensor 1420 and also have strong UV fluorescencecharacteristics. Thus such pesticide particles fit nicely within thedetection capabilities of the methods and apparatuses described above.Such pesticide particles may be distinguished from other types ofparticles based on morphological characteristics, color characteristicsunder visible light illumination as well as fluorescence colorcharacteristics under illumination by one or more UV light sources.

Detection and recognition of particles from pesticide sprays or dustingsare of interest for a number of purposes. In cases where pesticideparticles are a potential background for the detection of otherparticles of interest, it is desirable to recognize pesticide particlesso that they can be removed from the analysis of images containing otherparticles of interest. In other cases, the pesticide particles arethemselves of interest. For example, for farm worker safety, afterapplication of a dusting of fungicide, it may be desirable to determinewhen the fungicide has completely settled out of the ambient air andhence it is safe for a farm worker to re-enter the area. In anotherexample, it may be of interest for a remote farm manager to know whichfield was sprayed when with which pesticide. In some cases the farmmanager may appreciate receiving such information provided by a monitorin the field as well as through communications with farmworkers. In somecases the “farm workers” may be automatic robotic equipment.

Furthermore, it may be of interest to know the details of how thedensity of pesticide particles in ambient air decays with time. Forexample, it may be good to know if the amount of pesticide particles inthe air drops rapidly, perhaps because a sudden breeze blew thepesticide spray away from the intended field. Finally, we note that muchuseful information may be provided by correlations, or instead oradditionally, anti-correlations, between the detection of fungicide orpesticide particles and measured pathogenic fungal spores or otherpests.

Sooty Pollen Grains

The different color signatures of polycyclic aromatic hydrocarbons(PAHs) in soot particles and fluorescing molecules in biologicalparticles makes possible another technique. As pollen and sporeparticles are transported in air containing diesel exhaust pollution,their surfaces may make contact with, and adhere to, soot particles.This leads to techniques of using pollen grains being transported by thewind as probes of diesel particulate matter (DPM) in the air. Largerpollen grains may be best suited for this purpose as they are easilyimaged and recognized in an optical-microscope based devices.Furthermore, large pollen grains may more efficiently collect soot fromambient air as they respond to both settling from gravity andcentrifugal effects from air turbulence with larger relative velocitieswith respect to neighboring sub-micron soot particles.

Greater velocity with respect to small soot particles leads to morelarger pollen and spore particles encountering more small sootparticles. A sooty pollen grain, when captured and imaged, will have UVfluorescence with color signatures of biological molecules from itsinterior, but additional UV fluorescence due to diesel soot particles onits surface. The presence of surface diesel particles on the surface ofa biological particle may be determined from the color signatures ofdiesel particulate matter (DPM). Note that the species of pollen, andthe location of plants of the species within the local geography,provide clues regarding the path of the sooty pollen grain from itssource to the monitor, and hence clues regarding the location(s) wherethe pollen grain encountered soot particles.

Smoke & First Responders

The above described methods and apparatuses for collecting usefulinformation about diesel soot in ambient air also applies to soot andsmoke encountered by first responders to fire emergencies. Again,particle size distribution information and UV fluorescencecharacteristics may provide useful clues regarding the risk to humanhealth posed by smoke and soot in the ambient air. Also, on the theme ofmonitoring the health risks of ambient air, it may be of interest todetect of particles in exhaust from military use of ammunition andpropellants.

FIG. 52 is a simplified block diagram of a distributed computer network5200 that may be used in a specific embodiment of a system for airborneparticle collection, detection and recognition. Computer network 5200includes a number of client systems 5213, 5216, and 5219, and a serversystem 5222 coupled to a communication network 5224 via a plurality ofcommunication links 5228. There may be any number of clients and serversin a system. Communication network 5224 provides a mechanism forallowing the various components of distributed network 5200 tocommunicate and exchange information with each other.

Communication network 5224 may itself be comprised of manyinterconnected computer systems and communication links. Communicationlinks 5228 may be hardwire links, optical links, satellite or otherwireless communications links, wave propagation links, or any othermechanisms for communication of information. Various communicationprotocols may be used to facilitate communication between the varioussystems shown in FIG. 52. These communication protocols may includeTCP/IP, HTTP protocols, wireless application protocol (WAP),vendor-specific protocols, customized protocols, and others. While inone embodiment, communication network 5224 is the Internet, in otherembodiments, communication network 5224 may be any suitablecommunication network including a local area network (LAN), a wide areanetwork (WAN), a wireless network, an intranet, a private network, apublic network, a switched network, and combinations of these, and thelike.

Distributed computer network 5200 in FIG. 52 is merely illustrative ofan embodiment and is not intended to limit the scope of the embodimentas recited in the claims. One of ordinary skill in the art wouldrecognize other variations, modifications, and alternatives. Forexample, more than one server system 5222 may be connected tocommunication network 5224. As another example, a number of clientsystems 5213, 5216, and 5219 may be coupled to communication network5224 via an access provider (not shown) or via some other server system.

Client systems 5213, 5216, and 5219 enable users to access and queryinformation stored by server system 5222. In a specific embodiment, a“Web browser” application executing on a client system enables users toselect, access, retrieve, or query information stored by server system5222. Examples of web browsers include the Internet Explorer® and Edge®browser programs provided by Microsoft® Corporation, Chrome® browserprovided by Google®, and the Firefox® browser provided by Mozilla®Foundation, and others. In another specific embodiment, an iOS App or anAndroid® App on a client tablet enables users to select, access,retrieve, or query information stored by server system 5222. Access tothe system can be through a mobile application program or app that isseparate from a browser.

A computer-implemented or computer-executable version of the system maybe embodied using, stored on, or associated with computer-readablemedium or non-transitory computer-readable medium. A computer-readablemedium may include any medium that participates in providinginstructions to one or more processors for execution. Such a medium maytake many forms including, but not limited to, nonvolatile, volatile,and transmission media. Nonvolatile media includes, for example, flashmemory, or optical or magnetic disks. Volatile media includes static ordynamic memory, such as cache memory or RAM. Transmission media includescoaxial cables, copper wire, fiber optic lines, and wires arranged in abus. Transmission media can also take the form of electromagnetic, radiofrequency, acoustic, or light waves, such as those generated duringradio wave and infrared data communications.

For example, a binary, machine-executable version, of the software ofthe present system may be stored or reside in RAM or cache memory, or ona mass storage device. The source, executable code, or both of thesoftware may also be stored or reside on a mass storage device (e.g.,hard disk, magnetic disk, tape, or CD-ROM). As a further example, codemay be transmitted via wires, radio waves, or through a network such asthe Internet.

A client computer can be a smartphone, smartwatch, tablet computer,laptop, wearable device or computer (e.g., Google Glass), body-bornecomputer, or desktop.

FIG. 53 shows a system block diagram of computer system 5301. Computersystem 5301 includes monitor 5303, input device (e.g., keyboard,microphone, or camera) 5309, and mass storage devices 5317. Computersystem 5301 further includes subsystems such as central processor 5302,system memory 5304, input/output (I/O) controller 5306, display adapter5308, serial or universal serial bus (USB) port 5312, network interface5318, and speaker 5320. In an embodiment, a computer system includesadditional or fewer subsystems. For example, a computer system couldinclude more than one processor 5302 (i.e., a multiprocessor system) ora system may include a cache memory.

Arrows such as 5322 represent the system bus architecture of computersystem 5301. However, these arrows are illustrative of anyinterconnection scheme serving to link the subsystems. For example,speaker 5320 could be connected to the other subsystems through a portor have an internal direct connection to central processor 5302. Theprocessor may include multiple processors or a multicore processor,which may permit parallel processing of information. Computer system5301 shown in FIG. 53 is but an example of a suitable computer system.Other configurations of subsystems suitable for use will be readilyapparent to one of ordinary skill in the art.

Computer software products may be written in any of various suitableprogramming languages, such as C, C++, C#, Pascal, Fortran, Perl,Matlab® (from MathWorks), SAS, SPSS, JavaScript®, AJAX, Java®, SQL, andXQuery (a query language that is designed to process data from XML filesor any data source that can be viewed as XML, HTML, or both). Thecomputer software product may be an independent application with datainput and data display modules. Alternatively, the computer softwareproducts may be classes that may be instantiated as distributed objects.The computer software products may also be component software such asJava Beans® (from Oracle Corporation) or Enterprise Java Beans® (EJBfrom Oracle Corporation). In a specific embodiment, a computer programproduct is provided that stores instructions such as computer code toprogram a computer to perform any of the processes or techniquesdescribed.

An operating system for the system may be iOS by Apple®, Inc., Androidby Google®, one of the Microsoft Windows® family of operating systems(e.g., Windows NT®, Windows 2000®, Windows XP®, Windows XP® x64 Edition,Windows Vista®, Windows 7®, Windows CE®, Windows Mobile®, Windows 8,Windows 10), Linux, HP-UX, UNIX, Sun OS®, Solaris®, Mac OS X®, AlphaOS®, AIX, IRIX32, or IRIX64. Other operating systems may be used.Microsoft Windows® is a trademark of Microsoft® Corporation.

Furthermore, the computer may be connected to a network and mayinterface to other computers using this network. The network may be anintranet, internet, or the Internet, among others. The network may be awired network (e.g., using copper), telephone network, packet network,an optical network (e.g., using optical fiber), or a wireless network,or any combination of these. For example, data and other information maybe passed between the computer and components (or steps) of the systemusing a wireless network using a protocol such as Wi-Fi (IEEE standards802.11, 802.11a, 802.11b, 802.11e, 802.11g, 802.11i, and 802.11n, justto name a few examples). For example, signals from a computer may betransferred, at least in part, wirelessly to components or othercomputers.

In an embodiment, with a Web browser executing on a computer workstationsystem, a user accesses a system on the World Wide Web (WWW) through anetwork such as the Internet. The Web browser is used to download webpages or other content in various formats including HTML, XML, text,PDF, and postscript, and may be used to upload information to otherparts of the system. The Web browser may use uniform resourceidentifiers (URLs) to identify resources on the Web and hypertexttransfer protocol (HTTP) in transferring files on the Web.

In a specific embodiment, a method for determining a state of a fungalspore includes: directing a flow of air comprising the fungal spore to acollection cartridge; trapping the fungal spore within the collectioncartridge; illuminating the fungal spore in the collection cartridgewith light; while the fungal spore is illuminated with the light,capturing a first image of the fungal spore; analyzing the first imageto identify an outline of the fungal spore; illuminating the fungalspore in the collection cartridge with ultraviolet (UV) light; while thefungal spore is illuminated with the UV light, capturing a second imageof the fungal spore; measuring from the second image a degree offluorescence within the outline of the fungal spore; and based on thedegree of fluorescence, determining the state of the fungal spore.

Determining the state of the fungal spore may include determiningwhether the fungal spore is in a virulent state or a sterile state.Determining the state of the fungal spore may include comparing thedegree of fluorescence to a predetermined threshold value; if the degreeof fluorescence is above the predetermined value, determining that thefungal spore is in a first state; and if the degree of fluorescence isbelow the predetermined value, determining that the fungal spore is in asecond state, different from the first state.

In a specific embodiment, the method further includes inferringconcentrations of biomolecules of interest within the fungal spore bycorrelating a value of a pixel located at coordinates (x,y) on the imageto a concentration of a biomolecule of interest as being at thecoordinates (x,y) on the image; obtaining reference informationcomprising fluorescent properties of the biomolecules of interestassociated with known states of the fungal spore; obtaining camerasensor information of a camera sensor used to capture the second image,the camera sensor information comprising color sensitivitycharacteristics of the camera sensor; and processing the inferredconcentrations of the biomolecules of interest with the fluorescentproperties reference information and camera sensor information todetermine the state of the fungal spore.

The method may include storing in a log file a timestamp indicating whenthe fungal spore was trapped, and the determined state of the fungalspore, wherein the determined state comprises one of a virulent state ora sterile state.

In an embodiment, the collection cartridge includes a tape upon whichthe fungal spore is trapped and the method includes: after the trappingthe fungal spore within the collection cartridge, advancing the tapeupon which the fungal spore is trapped to a position underneath firstand second light sources, wherein the first light source comprises thelight, the second light source comprises the UV light, and whereinduring the illuminating the fungal spore with the light and illuminatingthe fungal spore with UV light, the tape remains in the same position.

In an embodiment, the method includes using an integrated camera sensorchip package to capturing the first image of the fungal spore, and thesecond image of the fungal spore, wherein the integrated camera sensorchip package comprises a light-sensing pixel sensor array, analog driveand readout circuitry, analog-to-digital conversion circuitry, digitalimage processing circuitry, and digital communications circuitry.

In another specific embodiment, there is a method for determining astate of a fungal spore comprising: defining a plurality of types offluorescent biomolecules of interest within the fungal spore; storing afirst plurality of predetermined color characteristics of fluorescentlight for each type of fluorescent biomolecules of interest, eachpredetermined color characteristic of the first plurality ofpredetermined color characteristics corresponding to excitement of arespective fluorescent biomolecule of interest under ultraviolet (UV)light having first spectral characteristics; storing a second pluralityof predetermined color characteristics of fluorescent light for eachtype of fluorescent biomolecules of interest, each predetermined colorcharacteristic of the second plurality of predetermined colorcharacteristics corresponding to excitement of a respective fluorescentbiomolecule of interest under UV light having second spectralcharacteristics, different from the first spectral characteristics;storing a third plurality of predetermined color characteristics offluorescent light for each type of fluorescent biomolecules of interest,each predetermined color characteristic of the third plurality ofpredetermined color characteristics corresponding to excitement of arespective fluorescent biomolecule of interest under UV light havingthird spectral characteristics, different from the first and secondspectral characteristics; directing a flow of air comprising the fungalspore to a collection cartridge; trapping the fungal spore within thecollection cartridge; illuminating the fungal spore in the collectioncartridge with light; while the fungal spore is illuminated with thelight, capturing a first two-dimensional color image of the fungalspore; analyzing the first two-dimensional color image to identify anoutline of the fungal spore, the outline of the fungal spore beingdefined by a set of image pixels receiving light from the fungal spore;illuminating the fungal spore in the collection cartridge with UV lightof the first spectral characteristics; while the fungal spore isilluminated with the UV light of the first spectral characteristics,capturing a second color image of the fungal spore; measuring from thesecond color image a degree and color of fluorescence for each pixelwithin the outline of the fungal spore; illuminating the fungal spore inthe collection cartridge with ultraviolet (UV) light of the secondspectral characteristics; while the fungal spore is illuminated with theUV light of the second spectral characteristics, capturing a third colorimage of the fungal spore; measuring from the third color image a degreeand color of fluorescence for each pixel within the outline of thefungal spore; illuminating the fungal spore in the collection cartridgewith UV light of the third spectral characteristics; while the fungalspore is illuminated with the UV light of the third spectralcharacteristics, capturing a fourth color image of the fungal spore;measuring from the fourth color image a degree and color of fluorescencefor each pixel within the outline of the fungal spore; based on themeasurements from the second, third, and fourth color images of degreeand color of fluorescence, estimating a concentration of each type offluorescent biomolecule of interest for each image pixel within theoutline of the fungal spore; generating two-dimensional images ofconcentrations of the fluorescent biomolecules of interest within theoutline of the fungal spore; and determining the state of the fungalspore from the estimated concentrations of each type of fluorescentbiomolecule of interest.

In an embodiment, the method includes using an integrated camera sensorchip package to capture the first two-dimensional color image of thefungal spore, and second, third, and fourth color images of the fungalspore, wherein the integrated camera sensor chip package comprises alight-sensing pixel sensor array, analog drive and readout circuitry,analog-to-digital conversion circuitry, digital image processingcircuitry, and digital communications circuitry.

In another specific embodiment, a method includes directing a flow ofair comprising a fungal spore to a collection cartridge; trapping thefungal spore on a tape medium of the collection cartridge; positioningthe fungal spore within a field of view of a camera sensor while thefungal spore remains trapped on the tape medium of the collectioncartridge; activating an ultraviolet (UV) light source to illuminate thetrapped fungal spore with UV light; opening a camera shutter associatedwith the camera sensor for a time period; while the trapped fungal sporeis illuminated with the UV light, allowing the camera sensor to collectlight emitted from the trapped fungal during a first portion of the timeperiod; after the first portion of the time period has elapsed,directing, during a second portion of the time period after the firstportion of the time period, a first burst of white light, originatingfrom a first position, towards the trapped fungal spore; directing,during the second portion of the time period, a second burst of whitelight, originating from a second position, different from the firstposition, towards the trapped fungal spore; after the second portion ofthe time period has elapsed, closing the camera shutter to generate animage; and analyzing the image to obtain a shape of the trapped fungalspore.

The method may further include directing, during the second portion ofthe time period, a third burst of white light, originating from a thirdposition, different from the first and second positions, towards thetrapped fungal spore.

In an embodiment, the first burst of white light is from a first lightemitting diode (LED) providing illumination corresponding to a firstcorner of the field of view, the second burst of white light is from asecond LED providing illumination corresponding to a second corner ofthe field of view, the third burst of white light is from a third LEDproviding illumination corresponding to a third corner of the field ofview, and the UV light source provides illumination corresponding to afourth corner of the field of view.

In an embodiment, the second portion of time period has a duration thatis less than a duration of the first portion of the time period. In anembodiment, a duration of the time period is about 15 seconds, aduration of the first portion of the time period is about 14 seconds,and a duration of the second portion of the time period is about 1second. In an embodiment, durations of each of the first and secondbursts of white light during the second portion of the time period areless than 1 second.

In an embodiment, the UV light source originates from a third position,different from the first and second positions, the first, second, andthird positions are arranged about the field of view of the camerasensor, and the first, second, and third positions are spaced120-degrees apart from each other.

In an embodiment, the method includes using an integrated camera sensorchip package to generate the image, wherein the integrated camera sensorchip package comprises a light-sensing pixel sensor array, analog driveand readout circuitry, analog-to-digital conversion circuitry, digitalimage processing circuitry, and digital communications circuitry.

The method may include after the analyzing the image to obtain a shapeof the trapped fungal spore, determining a state of the fungal spore.Determining a state of the fungal spore may include inferringconcentrations of biomolecules of interest within the fungal spore bycorrelating a value of a pixel located at coordinates (x,y) on the imageto a concentration of a biomolecule of interest as being at thecoordinates (x,y) on the image; obtaining reference informationcomprising fluorescent properties of the biomolecules of interestassociated with known states of the fungal spore; obtaining camerasensor information of the camera sensor, the camera sensor informationcomprising color sensitivity characteristics of the camera sensor; andprocessing the inferred concentrations of the biomolecules of interestwith the fluorescent properties reference information and camera sensorinformation to determine the state of the fungal spore.

In a specific embodiment, the first and second bursts of white light arefrom a single light source and the method includes: after the directinga first burst of white light, moving the single light source to thesecond position for the second burst of white light.

In the description above and throughout, numerous specific details areset forth in order to provide a thorough understanding of an embodimentof this disclosure. It will be evident, however, to one of ordinaryskill in the art, that an embodiment may be practiced without thesespecific details. In other instances, well-known structures and devicesare shown in block diagram form to facilitate explanation. Thedescription of the preferred embodiments is not intended to limit thescope of the claims appended hereto. Further, in the methods disclosedherein, various steps are disclosed illustrating some of the functionsof an embodiment. These steps are merely examples, and are not meant tobe limiting in any way. Other steps and functions may be contemplatedwithout departing from this disclosure or the scope of an embodiment.Other embodiments include systems and non-volatile media products thatexecute, embody or store processes that implement the methods describedabove.

What is claimed is:
 1. A method comprising: directing a flow of aircomprising a fungal spore to a collection cartridge; trapping the fungalspore on a tape medium of the collection cartridge; positioning thefungal spore within a field of view of a camera sensor while the fungalspore remains trapped on the tape medium of the collection cartridge;activating an ultraviolet (UV) light source to illuminate the trappedfungal spore with UV light; opening a camera shutter associated with thecamera sensor for a time period; while the trapped fungal spore isilluminated with the UV light, allowing the camera sensor to collectlight emitted from the trapped fungal during a first portion of the timeperiod; after the first portion of the time period has elapsed,directing, during a second portion of the time period after the firstportion of the time period, a first burst of white light, originatingfrom a first position, towards the trapped fungal spore; directing,during the second portion of the time period, a second burst of whitelight, originating from a second position, different from the firstposition, towards the trapped fungal spore; after the second portion ofthe time period has elapsed, closing the camera shutter to generate animage; and analyzing the image to obtain a shape of the trapped fungalspore.
 2. The method of claim 1 comprising: directing, during the secondportion of the time period, a third burst of white light, originatingfrom a third position, different from the first and second positions,towards the trapped fungal spore.
 3. The method of claim 2 wherein thefirst burst of white light is from a first light emitting diode (LED)providing illumination corresponding to a first corner of the field ofview, the second burst of white light is from a second LED providingillumination corresponding to a second corner of the field of view, thethird burst of white light is from a third LED providing illuminationcorresponding to a third corner of the field of view, and the UV lightsource provides illumination corresponding to a fourth corner of thefield of view.
 4. The method of claim 2 wherein the second portion oftime period has a duration that is less than a duration of the firstportion of the time period.
 5. The method of claim 2 wherein a durationof the time period is about 15 seconds, a duration of the first portionof the time period is about 14 seconds, and a duration of the secondportion of the time period is about 1 second.
 6. The method of claim 2wherein durations of each of the first and second bursts of white lightduring the second portion of the time period are less than 1 second. 7.The method of claim 1 wherein the UV light source originates from athird position, different from the first and second positions, andwherein the first, second, and third positions are arranged about thefield of view of the camera sensor, and wherein the first, second, andthird positions are spaced 120-degrees apart from each other.
 8. Themethod of claim 1 comprising: using an integrated camera sensor chippackage to generate the image, wherein the integrated camera sensor chippackage comprises a light-sensing pixel sensor array, analog drive andreadout circuitry, analog-to-digital conversion circuitry, digital imageprocessing circuitry, and digital communications circuitry.
 9. Themethod of claim 1 comprising: after the analyzing the image to obtain ashape of the trapped fungal spore, determining a state of the fungalspore.
 10. The method of claim 9 wherein the determining a state of thefungal spore comprises inferring concentrations of biomolecules ofinterest within the fungal spore by correlating a value of a pixellocated at coordinates (x,y) on the image to a concentration of abiomolecule of interest as being at the coordinates (x,y) on the image;obtaining reference information comprising fluorescent properties of thebiomolecules of interest associated with known states of the fungalspore; obtaining camera sensor information of the camera sensor, thecamera sensor information comprising color sensitivity characteristicsof the camera sensor; and processing the inferred concentrations of thebiomolecules of interest with the fluorescent properties referenceinformation and camera sensor information to determine the state of thefungal spore.
 11. The method of claim 1 wherein the first and secondbursts of white light are from a single light source and the methodcomprises: after the directing a first burst of white light, moving thesingle light source to the second position for the second burst of whitelight.